IRESite_Id:98 characteristics of HCV_type_1a IRES studied in pFGAL4-L270-HCV1 plasmid

IRESite: The database of experimentally verified IRES structures

The nucleic acid data:

IRESite Id: `98`	Version: `4`
Originaly submitted by: Tomáš Mašek	Submission date: `2005-12-15 00:00:00`
Reviewed by: Tomáš Mašek	Last change: `2008-07-04 12:37:02`

IRESite record type:

plasmid_with_promoter_and_putative_IRES_translationally_characterized

The shape of the nucleic acid molecule translated:
linear The quality of the mRNA/+RNA sequence:
both_UTRs_incomplete

The mRNA/+RNA description:

mRNA produced by tricistronic plasmid pFGAL4-L270 which comprises luciferase, L270 and Gal4 ORFs. L270 clone
was selected from pFGAL4::lambda library containing randomly cleaved phage lambda DNA for its low expression
of Gal4p in vivo. The full-length 5'-UTR of HCV genomic RNA together with the first 45 nt of a viral
polyprotein gene was inserted in frame into the reporter pFGAL4-L270 vector between the L270 and GAL4 ORFs,
thus making the N-terminal fusion of the first 15 amino acids residues from the HCV polyprotein with the yeast
transcriptional activator Gal4p.

The mRNA/+RNA sequence represented in the +DNA notation:

ATGGAAGACG CCAAAAACAT AAAGAAAGGC CCGGCGCCAT TCTATCCGCT GGAAGATGGA ACCGCTGGAG AGCAACTGCA TAAGGCTATG AAGAGATACG 
^1         ^11        ^21        ^31        ^41        ^51        ^61        ^71        ^81        ^91        

CCCTGGTTCC TGGAACAATT GCTTTTACAG ATGCACATAT CGAGGTGGAC ATCACTTACG CTGAGTACTT CGAAATGTCC GTTCGGTTGG CAGAAGCTAT 
^101       ^111       ^121       ^131       ^141       ^151       ^161       ^171       ^181       ^191       

GAAACGATAT GGGCTGAATA CAAATCACAG AATCGTCGTA TGCAGTGAAA ACTCTCTTCA ATTCTTTATG CCGGTGTTGG GCGCGTTATT TATCGGAGTT 
^201       ^211       ^221       ^231       ^241       ^251       ^261       ^271       ^281       ^291       

GCAGTTGCGC CCGCGAACGA CATTTATAAT GAACGTGAAT TGCTCAACAG TATGGGCATT TCGCAGCCTA CCGTGGTGTT CGTTTCCAAA AAGGGGTTGC 
^301       ^311       ^321       ^331       ^341       ^351       ^361       ^371       ^381       ^391       

AAAAAATTTT GAACGTGCAA AAAAAGCTCC CAATCATCCA AAAAATTATT ATCATGGATT CTAAAACGGA TTACCAGGGA TTTCAGTCGA TGTACACGTT 
^401       ^411       ^421       ^431       ^441       ^451       ^461       ^471       ^481       ^491       

CGTCACATCT CATCTACCTC CCGGTTTTAA TGAATACGAT TTTGTGCCAG AGTCCTTCGA TAGGGACAAG ACAATTGCAC TGATCATGAA CTCCTCTGGA 
^501       ^511       ^521       ^531       ^541       ^551       ^561       ^571       ^581       ^591       

TCTACTGGTC TGCCTAAAGG TGTCGCTCTG CCTCATAGAA CTGCCTGCGT GAGATTCTCG CATGCCAGAG ATCCTATTTT TGGCAATCAA ATCATTCCGG 
^601       ^611       ^621       ^631       ^641       ^651       ^661       ^671       ^681       ^691       

ATACTGCGAT TTTAAGTGTT GTTCCATTCC ATCACGGTTT TGGAATGTTT ACTACACTCG GATATTTGAT ATGTGGATTT CGAGTCGTCT TAATGTATAG 
^701       ^711       ^721       ^731       ^741       ^751       ^761       ^771       ^781       ^791       

ATTTGAAGAA GAGCTGTTTC TGAGGAGCCT TCAGGATTAC AAGATTCAAA GTGCGCTGCT GGTGCCAACC CTATTCTCCT TCTTCGCCAA AAGCACTCTG 
^801       ^811       ^821       ^831       ^841       ^851       ^861       ^871       ^881       ^891       

ATTGACAAAT ACGATTTATC TAATTTACAC GAAATTGCTT CTGGTGGCGC TCCCCTCTCT AAGGAAGTCG GGGAAGCGGT TGCCAAGAGG TTCCATCTGC 
^901       ^911       ^921       ^931       ^941       ^951       ^961       ^971       ^981       ^991       

CAGGTATCAG GCAAGGATAT GGGCTCACTG AGACTACATC AGCTATTCTG ATTACACCCG AGGGGGATGA TAAACCGGGC GCGGTCGGTA AAGTTGTTCC 
^1001      ^1011      ^1021      ^1031      ^1041      ^1051      ^1061      ^1071      ^1081      ^1091      

ATTTTTTGAA GCGAAGGTTG TGGATCTGGA TACCGGGAAA ACGCTGGGCG TTAATCAAAG AGGCGAACTG TGTGTGAGAG GTCCTATGAT TATGTCCGGT 
^1101      ^1111      ^1121      ^1131      ^1141      ^1151      ^1161      ^1171      ^1181      ^1191      

TATGTAAACA ATCCGGAAGC GACCAACGCC TTGATTGACA AGGATGGATG GCTACATTCT GGAGACATAG CTTACTGGGA CGAAGACGAA CACTTCTTCA 
^1201      ^1211      ^1221      ^1231      ^1241      ^1251      ^1261      ^1271      ^1281      ^1291      

TCGTTGACCG CCTGAAGTCT CTGATTAAGT ACAAAGGCTA TCAGGTGGCT CCCGCTGAAT TGGAATCCAT CTTGCTCCAA CACCCCAACA TCTTCGACGC 
^1301      ^1311      ^1321      ^1331      ^1341      ^1351      ^1361      ^1371      ^1381      ^1391      

AGGTGTCGCA GGTCTTCCCG ACGATGACGC CGGTGAACTT CCCGCCGCCG TTGTTGTTTT GGAGCACGGA AAGACGATGA CGGAAAAAGA GATCGTGGAT 
^1401      ^1411      ^1421      ^1431      ^1441      ^1451      ^1461      ^1471      ^1481      ^1491      

TACGTCGCCA GTCAAGTAAC AACCGCGAAA AAGTTGCGCG GAGGAGTTGT GTTTGTGGAC GAAGTACCGA AAGGTCTTAC CGGAAAACTC GACGCAAGAA 
^1501      ^1511      ^1521      ^1531      ^1541      ^1551      ^1561      ^1571      ^1581      ^1591      

AAATCAGAGA GATCCTCATA AAGGCCAAGA AGGGCGGAAA GATCGCCGTG TAATTCTAGC CTAGGGCCCA CAAGCTTGAA TTGTTTTATT TACTATCACA 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

AGATTAGTAG ATGTCGTTGG CGAAAAGTCA TTAGTTTTAT TCGATGAACC AGAGGTTCAT CTGCATCCAC CTTTGCTCTC TGCTTTTTTA CGAACATTAA 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

GCGACTTACT CGATGCACGC AATGGTGTAG CAATAATTCA GATCTCTCAC CTACCAAACA ATGCCCCCCT GCAAAAAATA AATTAACATC GCTCTTGAGC 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

AAAAAGGGTC CGGGAATTCG CCAGCCCCCT GATGGGGGCG ACACTCCACC ATGAATCACT CCCCTGTGAG GAACTACTGT CTTCACGCAG AAAGCGTCTA 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

GCCATGGCGT TAGTATGAGT GTCGTGCAGC CTCCAGGACC CCCCCTCCCG GGAGAGCCAT AGTGGTCTGC GGAACCGGTG AGTACACCGG AATTGCCAGG 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

ACGACCGGGT CCTTTCTTGG ATAAACCCGC TCAATGCCTG GAGATTTGGG CGTGCCCCCG CAAGACTGCT AGCCGAGTAG TGTTGGGTCG CGAAAGGCCT 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

TGTGGTACTG CCTGATAGGG TGCTTGCGAG TGCCCCGGGA GGTCTCGTAG ACCGTGCACC ATGAGCACGA ATCCTAAACC TCAAAGAAAA ACCAAACGTA 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

ACACGAATTC AAAAATGAAG CTACTGTCTT CTATCGAACA AGCATGCGAT ATTTGCCGAC TTAAAAAGCT CAAGTGCTCC AAAGAAAAAC CGAAGTGCGC 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

CAAGTGTCTG AAGAACAACT GGGAGTGTCG CTACTCTCCC AAAACCAAAA GGTCTCCGCT GACTAGGGCA CATCTGACAG AAGTGGAATC AAGGCTAGAA 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

AGACTGGAAC AGCTATTTCT ACTGATTTTT CCTCGAGAAG ACCTTGACAT GATTTTGAAA ATGGATTCTT TACAGGATAT AAAAGCATTG TTAACAGGAT 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

TATTTGTACA AGATAATGTG AATAAAGATG CCGTCACAGA TAGATTGGCT TCAGTGGAGA CTGATATGCC TCTAACATTG AGACAGCATA GAATAAGTGC 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

GACATCATCA TCGGAAGAGA GTAGTAACAA AGGTCAAAGA CAGTTGACTG TATCGATTGA CTCGGCAGCT CATCATGATA ACTCCACAAT TCCGTTGGAT 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

TTTATGCCCA GGGATGCTCT TCATGGATTT GATTGGTCTG AAGAGGATGA CATGTCGGAT GGCTTGCCCT TCCTGAAAAC GGACCCCAAC AATAATGGGT 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

TCTTTGGCGA CGGTTCTCTC TTATGTATTC TTCGATCTAT TGGCTTTAAA CCGGAAAATT ACACGAACTC TAACGTTAAC AGGCTCCCGA CCATGATTAC 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

GGATAGATAC ACGTTGGCTT CTAGATCCAC AACATCCCGT TTACTTCAAA GTTATCTCAA TAATTTTCAC CCCTACTGCC CTATCGTGCA CTCACCGACG 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

CTAATGATGT TGTATAATAA CCAGATTGAA ATCGCGTCGA AGGATCAATG GCAAATCCTT TTTAACTGCA TATTAGCCAT TGGAGCCTGG TGTATAGAGG 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

GGGAATCTAC TGATATAGAT GTTTTTTACT ATCAAAATGC TAAATCTCAT TTGACGAGCA AGGTCTTCGA GTCAGGTTCC ATAATTTTGG TGACAGCCCT 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

ACATCTTCTG TCGCGATATA CACAGTGGAG GCAGAAAACA AATACTAGCT ATAATTTTCA CAGCTTTTCC ATAAGAATGG CCATATCATT GGGCTTGAAT 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

AGGGACCTCC CCTCGTCCTT CAGTGATAGC AGCATTCTGG AACAAAGACG CCGAATTTGG TGGTCTGTCT ACTCTTGGGA GATCCAATTG TCCCTGCTTT 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

ATGGTCGATC CATCCAGCTT TCTCAGAATA CAATCTCCTT CCCTTCTTCT GTCGACGATG TGCAGCGTAC CACAACAGGT CCCACCATAT ATCATGGCAT 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

CATTGAAACA GCAAGGCTCT TACAAGTTTT CACAAAAATC TATGAACTAG ACAAAACAGT AACTGCAGAA AAAAGTCCTA TATGTGCAAA AAAATGCTTG 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

ATGATTTGTA ATGAGATTGA GGAGGTTTCG AGACAGGCAC CAAAGTTTTT ACAAATGGAT ATTTCCACCA CCGCTCTAAC CAATTTGTTG AAGGAACACC 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

CTTGGCTATC CTTTACAAGA TTCGAACTGA AGTGGAAACA GTTGTCTCTT ATCATTTATG TATTAAGAGA TTTTTTCACT AATTTTACCC AGAAAAAGTC 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

ACAACTAGAA CAGGATCAAA ATGATCATCA AAGTTATGAA GTTAAACGAT GCTCCATCAT GTTAAGCGAT GCAGCACAAA GAACTGTTAT GTCTGTAAGT 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

AGCTATATGG ACAATCATAA TGTCACCCCA TATTTTGCCT GGAATTGTTC TTATTACTTG TTCAATGCAG TCCTAGTACC CATAAAGACT CTACTCTCAA 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

ACTCAAAATC GAATGCTGAG AATAACGAGA CCGCACAATT ATTACAACAA ATTAACACTG TTCTGATGCT ATTAAAAAAA CTGGCCACTT TTAAAATCCA 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

GACTTGTGAA AAATACATTC AAGTACTGGA AGAGGTATGT GCGCCGTTTC TGTTATCACA GTGTGCAATC CCATTACCGC ATATCAGTTA TAACAATAGT 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

AATGGTAGCG CCATTAAAAA TATTGTCGGT TCTGCAACTA TCGCCCAATA CCCTACTCTT CCGGAGGAAA ATGTCAACAA TATCAGTGTT AAATATGTTT 
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371      ^4381      ^4391      

CTCCTGGCTC AGTAGGGCCT TCACCTGTGC CATTGAAATC AGGAGCAAGT TTCAGTGATC TAGTCAAGCT GTTATCTAAC CGTCCACCCT CTCGTAACTC 
^4401      ^4411      ^4421      ^4431      ^4441      ^4451      ^4461      ^4471      ^4481      ^4491      

TCCAGTGACA ATACCAAGAA GCACACCTTC GCATCGCTCA GTCACGCCTT TTCTAGGGCA ACAGCAACAG CTGCAATCAT TAGTGCCACT GACCCCGTCT 
^4501      ^4511      ^4521      ^4531      ^4541      ^4551      ^4561      ^4571      ^4581      ^4591      

GCTTTGTTTG GTGGCGCCAA TTTTAATCAA AGTGGGAATA TTGCTGATAG CTCATTGTCC TTCACTTTCA CTAACAGTAG CAACGGTCCG AACCTCATAA 
^4601      ^4611      ^4621      ^4631      ^4641      ^4651      ^4661      ^4671      ^4681      ^4691      

CAACTCAAAC AAATTCTCAA GCGCTTTCAC AACCAATTGC CTCCTCTAAC GTTCATGATA ACTTCATGAA TAATGAAATC ACGGCTAGTA AAATTGATGA 
^4701      ^4711      ^4721      ^4731      ^4741      ^4751      ^4761      ^4771      ^4781      ^4791      

TGGTAATAAT TCAAAACCAC TGTCACCTGG TTGGACGGAC CAAACTGCGT ATAACGCGTT TGGAATCACT ACAGGGATGT TTAATACCAC TACAATGGAT 
^4801      ^4811      ^4821      ^4831      ^4841      ^4851      ^4861      ^4871      ^4881      ^4891      

GATGTATATA ACTATCTATT CGATGATGAA GATACCCCAC CAAACCCAAA AAAAGAGTAA
^4901      ^4911      ^4921      ^4931      ^4941      ^4951

Credibility of mRNA sequence:

end-to-end_sequence_completely_same_as_in_the_experiment

The name of the plasmid:
pFGAL4-L270-HCV1 The name of the promoter used to express this mRNA:
TPI

The in vivo produced transcripts are heterogeneous (due to any of promoter?/splicing?/cleavage?/breakage?):

not tested

The in vivo produced heterogeneous transcripts occur due to alternative splicing:

not tested

A promoter reported in cDNA corresponding to IRES sequence:

not tested

Summary of possible issues when IRES cDNA is experimentally transcribed in vivo:

Summary of experiments studying integrity of the in vivo transcripts in a particular host:

Integrity (uniformity) of mRNA tested using Northern-blot:
unclear_result

Integrity (uniformity) of mRNA tested using RNase protection:
not_tested

Integrity (uniformity) of mRNA tested using 5'-RACE:
not_tested

Integrity (uniformity) of mRNA tested using primer extension :
not_tested

Integrity (uniformity) of mRNA tested using RT-PCR:
not_tested

Integrity (uniformity) of mRNA tested using real-time quantitative polymerase chain reaction (rtqPCR):
not_tested

Integrity (uniformity) of mRNA tested using RNAi:
not_tested

Integrity (uniformity) of mRNA tested using S1 nuclease mapping:
not_tested

Cryptic promoter presence was confirmed by expression from a promoter-less plasmid:
not_tested

Cryptic promoter presence was confirmed in an experimental setup involving inducible promoter:
not_tested

Integrity (uniformity) of mRNA molecules or possible promoter presence expressed in vivo was tested using another method, please specify in Remarks:
not_tested

The organism used:

Saccharomyces cerevisiae pJ69a

The abbreviated name of the donor gene or virus from which this IRES was excised and inserted into the plasmid:
HCV1a

The origin of IRES in the plasmid:

viral

The donor organism of the IRES segment:

Hepatitis C virus type 1a

The DNA sequence of the plasmid in (+) orientation annotated by its secondary structure:

GAATTAATTC ACCATGGAAG ACGCCAAAAA CATAAAGAAA GGCCCGGCGC CATTCTATCC GCTGGAAGAT GGAACCGCTG GAGAGCAACT GCATAAGGCT 
^1         ^11        ^21        ^31        ^41        ^51        ^61        ^71        ^81        ^91        

ATGAAGAGAT ACGCCCTGGT TCCTGGAACA ATTGCTTTTA CAGATGCACA TATCGAGGTG GACATCACTT ACGCTGAGTA CTTCGAAATG TCCGTTCGGT 
^101       ^111       ^121       ^131       ^141       ^151       ^161       ^171       ^181       ^191       

TGGCAGAAGC TATGAAACGA TATGGGCTGA ATACAAATCA CAGAATCGTC GTATGCAGTG AAAACTCTCT TCAATTCTTT ATGCCGGTGT TGGGCGCGTT 
^201       ^211       ^221       ^231       ^241       ^251       ^261       ^271       ^281       ^291       

ATTTATCGGA GTTGCAGTTG CGCCCGCGAA CGACATTTAT AATGAACGTG AATTGCTCAA CAGTATGGGC ATTTCGCAGC CTACCGTGGT GTTCGTTTCC 
^301       ^311       ^321       ^331       ^341       ^351       ^361       ^371       ^381       ^391       

AAAAAGGGGT TGCAAAAAAT TTTGAACGTG CAAAAAAAGC TCCCAATCAT CCAAAAAATT ATTATCATGG ATTCTAAAAC GGATTACCAG GGATTTCAGT 
^401       ^411       ^421       ^431       ^441       ^451       ^461       ^471       ^481       ^491       

CGATGTACAC GTTCGTCACA TCTCATCTAC CTCCCGGTTT TAATGAATAC GATTTTGTGC CAGAGTCCTT CGATAGGGAC AAGACAATTG CACTGATCAT 
^501       ^511       ^521       ^531       ^541       ^551       ^561       ^571       ^581       ^591       

GAACTCCTCT GGATCTACTG GTCTGCCTAA AGGTGTCGCT CTGCCTCATA GAACTGCCTG CGTGAGATTC TCGCATGCCA GAGATCCTAT TTTTGGCAAT 
^601       ^611       ^621       ^631       ^641       ^651       ^661       ^671       ^681       ^691       

CAAATCATTC CGGATACTGC GATTTTAAGT GTTGTTCCAT TCCATCACGG TTTTGGAATG TTTACTACAC TCGGATATTT GATATGTGGA TTTCGAGTCG 
^701       ^711       ^721       ^731       ^741       ^751       ^761       ^771       ^781       ^791       

TCTTAATGTA TAGATTTGAA GAAGAGCTGT TTCTGAGGAG CCTTCAGGAT TACAAGATTC AAAGTGCGCT GCTGGTGCCA ACCCTATTCT CCTTCTTCGC 
^801       ^811       ^821       ^831       ^841       ^851       ^861       ^871       ^881       ^891       

CAAAAGCACT CTGATTGACA AATACGATTT ATCTAATTTA CACGAAATTG CTTCTGGTGG CGCTCCCCTC TCTAAGGAAG TCGGGGAAGC GGTTGCCAAG 
^901       ^911       ^921       ^931       ^941       ^951       ^961       ^971       ^981       ^991       

AGGTTCCATC TGCCAGGTAT CAGGCAAGGA TATGGGCTCA CTGAGACTAC ATCAGCTATT CTGATTACAC CCGAGGGGGA TGATAAACCG GGCGCGGTCG 
^1001      ^1011      ^1021      ^1031      ^1041      ^1051      ^1061      ^1071      ^1081      ^1091      

GTAAAGTTGT TCCATTTTTT GAAGCGAAGG TTGTGGATCT GGATACCGGG AAAACGCTGG GCGTTAATCA AAGAGGCGAA CTGTGTGTGA GAGGTCCTAT 
^1101      ^1111      ^1121      ^1131      ^1141      ^1151      ^1161      ^1171      ^1181      ^1191      

GATTATGTCC GGTTATGTAA ACAATCCGGA AGCGACCAAC GCCTTGATTG ACAAGGATGG ATGGCTACAT TCTGGAGACA TAGCTTACTG GGACGAAGAC 
^1201      ^1211      ^1221      ^1231      ^1241      ^1251      ^1261      ^1271      ^1281      ^1291      

GAACACTTCT TCATCGTTGA CCGCCTGAAG TCTCTGATTA AGTACAAAGG CTATCAGGTG GCTCCCGCTG AATTGGAATC CATCTTGCTC CAACACCCCA 
^1301      ^1311      ^1321      ^1331      ^1341      ^1351      ^1361      ^1371      ^1381      ^1391      

ACATCTTCGA CGCAGGTGTC GCAGGTCTTC CCGACGATGA CGCCGGTGAA CTTCCCGCCG CCGTTGTTGT TTTGGAGCAC GGAAAGACGA TGACGGAAAA 
^1401      ^1411      ^1421      ^1431      ^1441      ^1451      ^1461      ^1471      ^1481      ^1491      

AGAGATCGTG GATTACGTCG CCAGTCAAGT AACAACCGCG AAAAAGTTGC GCGGAGGAGT TGTGTTTGTG GACGAAGTAC CGAAAGGTCT TACCGGAAAA 
^1501      ^1511      ^1521      ^1531      ^1541      ^1551      ^1561      ^1571      ^1581      ^1591      

CTCGACGCAA GAAAAATCAG AGAGATCCTC ATAAAGGCCA AGAAGGGCGG AAAGATCGCC GTGTAATTCT AGCCTAGGGC CCACAAGCTT GAATTGTTTT 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

ATTTACTATC ACAAGATTAG TAGATGTCGT TGGCGAAAAG TCATTAGTTT TATTCGATGA ACCAGAGGTT CATCTGCATC CACCTTTGCT CTCTGCTTTT 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

TTACGAACAT TAAGCGACTT ACTCGATGCA CGCAATGGTG TAGCAATAAT TCAGATCTCT CACCTACCAA ACAATGCCCC CCTGCAAAAA ATAAATTAAC 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

ATCGCTCTTG AGCAAAAAGG GTCCGGGAAT TCGCCAGCCC CCTGATGGGG GCGACACTCC ACCATGAATC ACTCCCCTGT GAGGAACTAC TGTCTTCACG 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

CAGAAAGCGT CTAGCCATGG CGTTAGTATG AGTGTCGTGC AGCCTCCAGG ACCCCCCCTC CCGGGAGAGC CATAGTGGTC TGCGGAACCG GTGAGTACAC 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

CGGAATTGCC AGGACGACCG GGTCCTTTCT TGGATAAACC CGCTCAATGC CTGGAGATTT GGGCGTGCCC CCGCAAGACT GCTAGCCGAG TAGTGTTGGG 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

TCGCGAAAGG CCTTGTGGTA CTGCCTGATA GGGTGCTTGC GAGTGCCCCG GGAGGTCTCG TAGACCGTGC ACCATGAGCA CGAATCCTAA ACCTCAAAGA 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

AAAACCAAAC GTAACACGAA TTCAAAAATG AAGCTACTGT CTTCTATCGA ACAAGCATGC GATATTTGCC GACTTAAAAA GCTCAAGTGC TCCAAAGAAA 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

AACCGAAGTG CGCCAAGTGT CTGAAGAACA ACTGGGAGTG TCGCTACTCT CCCAAAACCA AAAGGTCTCC GCTGACTAGG GCACATCTGA CAGAAGTGGA 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

ATCAAGGCTA GAAAGACTGG AACAGCTATT TCTACTGATT TTTCCTCGAG AAGACCTTGA CATGATTTTG AAAATGGATT CTTTACAGGA TATAAAAGCA 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

TTGTTAACAG GATTATTTGT ACAAGATAAT GTGAATAAAG ATGCCGTCAC AGATAGATTG GCTTCAGTGG AGACTGATAT GCCTCTAACA TTGAGACAGC 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

ATAGAATAAG TGCGACATCA TCATCGGAAG AGAGTAGTAA CAAAGGTCAA AGACAGTTGA CTGTATCGAT TGACTCGGCA GCTCATCATG ATAACTCCAC 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

AATTCCGTTG GATTTTATGC CCAGGGATGC TCTTCATGGA TTTGATTGGT CTGAAGAGGA TGACATGTCG GATGGCTTGC CCTTCCTGAA AACGGACCCC 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

AACAATAATG GGTTCTTTGG CGACGGTTCT CTCTTATGTA TTCTTCGATC TATTGGCTTT AAACCGGAAA ATTACACGAA CTCTAACGTT AACAGGCTCC 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

CGACCATGAT TACGGATAGA TACACGTTGG CTTCTAGATC CACAACATCC CGTTTACTTC AAAGTTATCT CAATAATTTT CACCCCTACT GCCCTATCGT 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

GCACTCACCG ACGCTAATGA TGTTGTATAA TAACCAGATT GAAATCGCGT CGAAGGATCA ATGGCAAATC CTTTTTAACT GCATATTAGC CATTGGAGCC 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

TGGTGTATAG AGGGGGAATC TACTGATATA GATGTTTTTT ACTATCAAAA TGCTAAATCT CATTTGACGA GCAAGGTCTT CGAGTCAGGT TCCATAATTT 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

TGGTGACAGC CCTACATCTT CTGTCGCGAT ATACACAGTG GAGGCAGAAA ACAAATACTA GCTATAATTT TCACAGCTTT TCCATAAGAA TGGCCATATC 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

ATTGGGCTTG AATAGGGACC TCCCCTCGTC CTTCAGTGAT AGCAGCATTC TGGAACAAAG ACGCCGAATT TGGTGGTCTG TCTACTCTTG GGAGATCCAA 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

TTGTCCCTGC TTTATGGTCG ATCCATCCAG CTTTCTCAGA ATACAATCTC CTTCCCTTCT TCTGTCGACG ATGTGCAGCG TACCACAACA GGTCCCACCA 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

TATATCATGG CATCATTGAA ACAGCAAGGC TCTTACAAGT TTTCACAAAA ATCTATGAAC TAGACAAAAC AGTAACTGCA GAAAAAAGTC CTATATGTGC 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

AAAAAAATGC TTGATGATTT GTAATGAGAT TGAGGAGGTT TCGAGACAGG CACCAAAGTT TTTACAAATG GATATTTCCA CCACCGCTCT AACCAATTTG 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

TTGAAGGAAC ACCCTTGGCT ATCCTTTACA AGATTCGAAC TGAAGTGGAA ACAGTTGTCT CTTATCATTT ATGTATTAAG AGATTTTTTC ACTAATTTTA 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

CCCAGAAAAA GTCACAACTA GAACAGGATC AAAATGATCA TCAAAGTTAT GAAGTTAAAC GATGCTCCAT CATGTTAAGC GATGCAGCAC AAAGAACTGT 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

TATGTCTGTA AGTAGCTATA TGGACAATCA TAATGTCACC CCATATTTTG CCTGGAATTG TTCTTATTAC TTGTTCAATG CAGTCCTAGT ACCCATAAAG 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

ACTCTACTCT CAAACTCAAA ATCGAATGCT GAGAATAACG AGACCGCACA ATTATTACAA CAAATTAACA CTGTTCTGAT GCTATTAAAA AAACTGGCCA 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

CTTTTAAAAT CCAGACTTGT GAAAAATACA TTCAAGTACT GGAAGAGGTA TGTGCGCCGT TTCTGTTATC ACAGTGTGCA ATCCCATTAC CGCATATCAG 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

TTATAACAAT AGTAATGGTA GCGCCATTAA AAATATTGTC GGTTCTGCAA CTATCGCCCA ATACCCTACT CTTCCGGAGG AAAATGTCAA CAATATCAGT 
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371      ^4381      ^4391      

GTTAAATATG TTTCTCCTGG CTCAGTAGGG CCTTCACCTG TGCCATTGAA ATCAGGAGCA AGTTTCAGTG ATCTAGTCAA GCTGTTATCT AACCGTCCAC 
^4401      ^4411      ^4421      ^4431      ^4441      ^4451      ^4461      ^4471      ^4481      ^4491      

CCTCTCGTAA CTCTCCAGTG ACAATACCAA GAAGCACACC TTCGCATCGC TCAGTCACGC CTTTTCTAGG GCAACAGCAA CAGCTGCAAT CATTAGTGCC 
^4501      ^4511      ^4521      ^4531      ^4541      ^4551      ^4561      ^4571      ^4581      ^4591      

ACTGACCCCG TCTGCTTTGT TTGGTGGCGC CAATTTTAAT CAAAGTGGGA ATATTGCTGA TAGCTCATTG TCCTTCACTT TCACTAACAG TAGCAACGGT 
^4601      ^4611      ^4621      ^4631      ^4641      ^4651      ^4661      ^4671      ^4681      ^4691      

CCGAACCTCA TAACAACTCA AACAAATTCT CAAGCGCTTT CACAACCAAT TGCCTCCTCT AACGTTCATG ATAACTTCAT GAATAATGAA ATCACGGCTA 
^4701      ^4711      ^4721      ^4731      ^4741      ^4751      ^4761      ^4771      ^4781      ^4791      

GTAAAATTGA TGATGGTAAT AATTCAAAAC CACTGTCACC TGGTTGGACG GACCAAACTG CGTATAACGC GTTTGGAATC ACTACAGGGA TGTTTAATAC 
^4801      ^4811      ^4821      ^4831      ^4841      ^4851      ^4861      ^4871      ^4881      ^4891      

CACTACAATG GATGATGTAT ATAACTATCT ATTCGATGAT GAAGATACCC CACCAAACCC AAAAAAAGAG TAAGAGCTCA GCTAGCTAAC TGAATAAGGA 
^4901      ^4911      ^4921      ^4931      ^4941      ^4951      ^4961      ^4971      ^4981      ^4991      

ACAATGAACG TTTTTCCTTT CTCTTGTTCC TAGTATTAAT GACTGACCGA TACATCCCTT TTTTTTTTTG TCTTTGTCTA GCTCCAATTC GCCCTATAGT 
^5001      ^5011      ^5021      ^5031      ^5041      ^5051      ^5061      ^5071      ^5081      ^5091      

GAGTCGTATT ACAATTCACT GGCCGTCGTT TTACAACGTC GTGACTGGGA AAACCCTGGC GTTACCCAAC TTAATCGCCT TGCAGCACAT CCCCCTTTCG 
^5101      ^5111      ^5121      ^5131      ^5141      ^5151      ^5161      ^5171      ^5181      ^5191      

CCAGCTGGCG TAATAGCGAA GAGGCCCGCA CCGATCGCCC TTCCCAACAG TTGCGCAGCC TGAATGGCGA ATGGACGCGC CCTGTAGCGG CGCATTAAGC 
^5201      ^5211      ^5221      ^5231      ^5241      ^5251      ^5261      ^5271      ^5281      ^5291      

GCGGCGGGTG TGGTGGTTAC GCGCAGCGTG ACCGCTACAC TTGCCAGCGC CCTAGCGCCC GCTCCTTTCG CTTTCTTCCC TTCCTTTCTC GCCACGTTCG 
^5301      ^5311      ^5321      ^5331      ^5341      ^5351      ^5361      ^5371      ^5381      ^5391      

CCGGCTTTCC CCGTCAAGCT CTAAATCGGG GGCTCCCTTT AGGGTTCCGA TTTAGTGCTT TACGGCACCT CGACCCCAAA AAACTTGATT AGGGTGATGG 
^5401      ^5411      ^5421      ^5431      ^5441      ^5451      ^5461      ^5471      ^5481      ^5491      

TTCACGTAGT GGGCCATCGC CCTGATAGAC GGTTTTTCGC CCTTTGACGT TGGAGTCCAC GTTCTTTAAT AGTGGACTCT TGTTCCAAAC TGGAACAACA 
^5501      ^5511      ^5521      ^5531      ^5541      ^5551      ^5561      ^5571      ^5581      ^5591      

CTCAACCCTA TCTCGGTCTA TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT TGGTTAAAAA ATGAGCTGAT TTAACAAAAA TTTAACGCGA 
^5601      ^5611      ^5621      ^5631      ^5641      ^5651      ^5661      ^5671      ^5681      ^5691      

ATTTTAACAA AATATTAACG CTTACAATTT CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT TCACACCGCA TAGGGTAATA ACTGATATAA 
^5701      ^5711      ^5721      ^5731      ^5741      ^5751      ^5761      ^5771      ^5781      ^5791      

TTAAATTGAA GCTCTAATTT GTGAGTTTAG TATACATGCA TTTACTTATA ATACAGTTTT TTAGTTTTGC TGGCCGCATC TTCTCAAATA TGCTTCCCAG 
^5801      ^5811      ^5821      ^5831      ^5841      ^5851      ^5861      ^5871      ^5881      ^5891      

CCTGCTTTTC TGTAACGTTC ACCCTCTACC TTAGCATCCC TTCCCTTTGC AAATAGTCCT CTTCCAACAA TAATAATGTC AGATCCTGTA GAGACCACAT 
^5901      ^5911      ^5921      ^5931      ^5941      ^5951      ^5961      ^5971      ^5981      ^5991      

CATCCACGGT TCTATACTGT TGACCCAATG CGTCTCCCTT GTCATCTAAA CCCACACCGG GTGTCATAAT CAACCAATCG TAACCTTCAT CTCTTCCACC 
^6001      ^6011      ^6021      ^6031      ^6041      ^6051      ^6061      ^6071      ^6081      ^6091      

CATGTCTCTT TGAGCAATAA AGCCGATAAC AAAATCTTTG TCGCTCTTCG CAATGTCAAC AGTACCCTTA GTATATTCTC CAGTAGATAG GGAGCCCTTG 
^6101      ^6111      ^6121      ^6131      ^6141      ^6151      ^6161      ^6171      ^6181      ^6191      

CATGACAATT CTGCTAACAT CAAAAGGCCT CTAGGTTCCT TTGTTACTTC TTCTGCCGCC TGCTTCAAAC CGCTAACAAT ACCTGGCCCC ACCACACCGT 
^6201      ^6211      ^6221      ^6231      ^6241      ^6251      ^6261      ^6271      ^6281      ^6291      

GTGCATTCGT AATGTCTGCC CATTCTGCTA TTCTGTATAC ACCCGCAGAG TACTGCAATT TGACTGTATT ACCAATGTCA GCAAATTTTC TGTCTTCGAA 
^6301      ^6311      ^6321      ^6331      ^6341      ^6351      ^6361      ^6371      ^6381      ^6391      

GAGTAAAAAA TTGTACTTGG CGGATAATGC CTTTAGCGGC TTAACTGTGC CCTCCATCGA AAAATCAGTC AATATATCCA CATGTGTTTT TAGTAAACAA 
^6401      ^6411      ^6421      ^6431      ^6441      ^6451      ^6461      ^6471      ^6481      ^6491      

ATTTTGGGAC CTAATGCTTC AACTAACTCC AGTAATTCCT TGGTGGTACG AACATCCAAT GAAGCACACA AGTTTGTTTG CTTTTCGTGC ATGATATTAA 
^6501      ^6511      ^6521      ^6531      ^6541      ^6551      ^6561      ^6571      ^6581      ^6591      

ATAGCTTGGC AGCAACAGGA CTAGGATGAG TAGCAGCACG TTCCTTATAT GTAGCTTTCG ACATGATTTA TCTTCGTTTC CTGCAGGTTT TTGTTCTGTG 
^6601      ^6611      ^6621      ^6631      ^6641      ^6651      ^6661      ^6671      ^6681      ^6691      

CAGTTGGGTT AAGAATACTG GGCAATTTCA TGTTTCTTCA ACACTACATA TGCGTATATA TACCAATCTA AGTCTGTGCT CCTTCCTTCG TTCTTCCTTC 
^6701      ^6711      ^6721      ^6731      ^6741      ^6751      ^6761      ^6771      ^6781      ^6791      

TGTTCGGAGA TTACCGAATC AAAAAAATTT CAAGGAAACC GAAATCAAAA AAAAGAATAA AAAAAAAATG ATGAATTGAA AAGGTGGTAT GGTGCACTCT 
^6801      ^6811      ^6821      ^6831      ^6841      ^6851      ^6861      ^6871      ^6881      ^6891      

CAGTACAATC TGCTCTGATG CCGCATAGTT AAGCCAGCCC CGACACCCGC CAACACCCGC TGACGCGCCC TGACGGGCTT GTCTGCTCCC GGCATCCGCT 
^6901      ^6911      ^6921      ^6931      ^6941      ^6951      ^6961      ^6971      ^6981      ^6991      

TACAGACAAG CTGTGACCGT CTCCGGGAGC TGCATGTGTC AGAGGTTTTC ACCGTCATCA CCGAAACGCG CGAGACGAAA GGGCCTCGTG ATACGCCTAT 
^7001      ^7011      ^7021      ^7031      ^7041      ^7051      ^7061      ^7071      ^7081      ^7091      

TTTTATAGGT TAATGTCATG ATAATAATGG TTTCTTAGAC GTGCGGCCGC TCTAGAACTA GTGGATCAAT TCCACGGACT ATAGACTATA CTAGTATACT 
^7101      ^7111      ^7121      ^7131      ^7141      ^7151      ^7161      ^7171      ^7181      ^7191      

CCGTCTACTG TACGATACAC TTCCGCTCAG GTCCTTGTCC TTTAACGAGG CCTTACCACT CTTTTGTTAC TCTATTGATC CAGCTCAGCA AAGGCAGTGT 
^7201      ^7211      ^7221      ^7231      ^7241      ^7251      ^7261      ^7271      ^7281      ^7291      

GATCTAAGAT TCTATCTTCG CGATGTAGTA AAACTAGCTA GACCGAGAAA GAGACTAGAA ATGCAAAAGG CACTTCTACA ATGGCTGCCA TCATTATTAT 
^7301      ^7311      ^7321      ^7331      ^7341      ^7351      ^7361      ^7371      ^7381      ^7391      

CCGATGTGAC GCTGCAGCTT CTCAATGATA TTCGAATACG CTTTGAGGAG ATACAGCCTA ATATCCGACA AACTGTTTTA CAGATTTACG ATCGTACTTG 
^7401      ^7411      ^7421      ^7431      ^7441      ^7451      ^7461      ^7471      ^7481      ^7491      

TTACCCATCA TTGAATTTTG AACATCCGAA CCTGGGAGTT TTCCCTGAAA CAGATAGTAT ATTTGAACCT GTATAATAAT ATATAGTCTA GCGCTTTACG 
^7501      ^7511      ^7521      ^7531      ^7541      ^7551      ^7561      ^7571      ^7581      ^7591      

GAAGACAATG TATGTATTTC GGTTCCTGGA GAAACTATTG CATCTATTGC ATAGGTAATC TTGCACGTCG CATCCCCGGT TCATTTTCTG CGTTTCCATC 
^7601      ^7611      ^7621      ^7631      ^7641      ^7651      ^7661      ^7671      ^7681      ^7691      

TTGCACTTCA ATAGCATATC TTTGTTAACG AAGCATCTGT GCTTCATTTT GTAGAACAAA AATGCAACGC GAGAGCGCTA ATTTTTCAAA CAAAGAATCT 
^7701      ^7711      ^7721      ^7731      ^7741      ^7751      ^7761      ^7771      ^7781      ^7791      

GAGCTGCATT TTTACAGAAC AGAAATGCAA CGCGAAAGCG CTATTTTACC AACGAAGAAT CTGTGCTTCA TTTTTGTAAA ACAAAAATGC AACGCGAGAG 
^7801      ^7811      ^7821      ^7831      ^7841      ^7851      ^7861      ^7871      ^7881      ^7891      

CGCTAATTTT TCAAACAAAG AATCTGAGCT GCATTTTTAC AGAACAGAAA TGCAACGCGA GAGCGCTATT TTACCAACAA AGAATCTATA CTTCTTTTTT 
^7901      ^7911      ^7921      ^7931      ^7941      ^7951      ^7961      ^7971      ^7981      ^7991      

GTTCTACAAA AATGCATCCC GAGAGCGCTA TTTTTCTAAC AAAGCATCTT AGATTACTTT TTTTCTCCTT TGTGCGCTCT ATAATGCAGT CTCTTGATAA 
^8001      ^8011      ^8021      ^8031      ^8041      ^8051      ^8061      ^8071      ^8081      ^8091      

CTTTTTGCAC TGTAGGTCCG TTAAGGTTAG AAGAAGGCTA CTTTGGTGTC TATTTTCTCT TCCATAAAAA AAGCCTGACT CCACTTCCCG CGTTTACTGA 
^8101      ^8111      ^8121      ^8131      ^8141      ^8151      ^8161      ^8171      ^8181      ^8191      

TTACTAGCGA AGCTGCGGGT GCATTTTTTC AAGATAAAGG CATCCCCGAT TATATTCTAT ACCGATGTGG ATTGCGCATA CTTTGTGAAC AGAAAGTGAT 
^8201      ^8211      ^8221      ^8231      ^8241      ^8251      ^8261      ^8271      ^8281      ^8291      

AGCGTTGATG ATTCTTCATT GGTCAGAAAA TTATGAACGG TTTCTTCTAT TTTGTCTCTA TATACTACGT ATAGGAAATG TTTACATTTT CGTATTGTTT 
^8301      ^8311      ^8321      ^8331      ^8341      ^8351      ^8361      ^8371      ^8381      ^8391      

TCGATTCACT CTATGAATAG TTCTTACTAC AATTTTTTTG TCTAAAGAGT AATACTAGAG ATAAACATAA AAAATGTAGA GGTCGAGTTT AGATGCAAGT 
^8401      ^8411      ^8421      ^8431      ^8441      ^8451      ^8461      ^8471      ^8481      ^8491      

TCAAGGAGCG AAAGGTGGAT GGGTAGGTTA TATAGGGATA TAGCACAGAG ATATATAGCA AAGAGATACT TTTGAGCAAT GTTTGTGGAA GCGGTATTCG 
^8501      ^8511      ^8521      ^8531      ^8541      ^8551      ^8561      ^8571      ^8581      ^8591      

CAATATTTTA GTAGCTCGTT ACAGTCCGGT GCGTTTTTGG TTTTTTGAAA GTGCGTCTTC AGAGCGCTTT TGGTTTTCAA AAGCGCTCTG AAGTTCCTAT 
^8601      ^8611      ^8621      ^8631      ^8641      ^8651      ^8661      ^8671      ^8681      ^8691      

ACTTTCTAGA GAATAGGAAC TTCGGAATAG GAACTTCAAA GCGTTTCCGA AAACGAGCGC TTCCGAAAAT GCAACGCGAG CTGCGCACAT ACAGCTCACT 
^8701      ^8711      ^8721      ^8731      ^8741      ^8751      ^8761      ^8771      ^8781      ^8791      

GTTCACGTCG CACCTATATC TGCGTGTTGC CTGTATATAT ATATACATGA GAAGAACGGC ATAGTGCGTG TTTATGCTTA AATGCGTACT TATATGCGTC 
^8801      ^8811      ^8821      ^8831      ^8841      ^8851      ^8861      ^8871      ^8881      ^8891      

TATTTATGTA GGATGAAAGG TAGTCTAGTA CCTCCTGTGA TATTATCCCA TTCCATGCGG GGTATCGTAT GCTTCCTTCA GCACTACCCT TTAGCTGTTC 
^8901      ^8911      ^8921      ^8931      ^8941      ^8951      ^8961      ^8971      ^8981      ^8991      

TATATGCTGC CACTCCTCAA TTGGATTAGT CTCATCCTTC AATGCTATCA TTTCCTTTGA TATTGGATCA TATGCATAGT ACCGAGAAAC TAGTGCGAAG 
^9001      ^9011      ^9021      ^9031      ^9041      ^9051      ^9061      ^9071      ^9081      ^9091      

TAGTGATCAG GTATTGCTGT TATCTGATGA GTATACGTTG TCCTGGCCAC GGCAGAAGCA CGCTTATCGC TCCAATTTCC CACAACATTA GTCAACTCCG 
^9101      ^9111      ^9121      ^9131      ^9141      ^9151      ^9161      ^9171      ^9181      ^9191      

TTAGGCCCTT CATTGAAAGA AATGAGGTCA TCAAATGTCT TCCAATGTGA GATTTTGGGC CATTTTTTAT AGCAAAGATT GAATAAGGCG CATTTTTCTT 
^9201      ^9211      ^9221      ^9231      ^9241      ^9251      ^9261      ^9271      ^9281      ^9291      

CAAAGCTGCG GCCGCACTCT CACTAGTACG TCAGGTGGCA CTTTTCGGGG AAATGTGCGC GGAACCCCTA TTTGTTTATT TTTCTAAATA CATTCAAATA 
^9301      ^9311      ^9321      ^9331      ^9341      ^9351      ^9361      ^9371      ^9381      ^9391      

TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA ATAATATTGA AAAAGGAAGA GTATGAGTAT TCAACATTTC CGTGTCGCCC TTATTCCCTT 
^9401      ^9411      ^9421      ^9431      ^9441      ^9451      ^9461      ^9471      ^9481      ^9491      

TTTTGCGGCA TTTTGCCTTC CTGTTTTTGC TCACCCAGAA ACGCTGGTGA AAGTAAAAGA TGCTGAAGAT CAGTTGGGTG CACGAGTGGG TTACATCGAA 
^9501      ^9511      ^9521      ^9531      ^9541      ^9551      ^9561      ^9571      ^9581      ^9591      

CTGGATCTCA ACAGCGGTAA GATCCTTGAG AGTTTTCGCC CCGAAGAACG TTTTCCAATG ATGAGCACTT TTAAAGTTCT GCTATGTGGC GCGGTATTAT 
^9601      ^9611      ^9621      ^9631      ^9641      ^9651      ^9661      ^9671      ^9681      ^9691      

CCCGTATTGA CGCCGGGCAA GAGCAACTCG GTCGCCGCAT ACACTATTCT CAGAATGACT TGGTTGAGTA CTCACCAGTC ACAGAAAAGC ATCTTACGGA 
^9701      ^9711      ^9721      ^9731      ^9741      ^9751      ^9761      ^9771      ^9781      ^9791      

TGGCATGACA GTAAGAGAAT TATGCAGTGC TGCCATAACC ATGAGTGATA ACACTGCGGC CAACTTACTT CTGACAACGA TCGGAGGACC GAAGGAGCTA 
^9801      ^9811      ^9821      ^9831      ^9841      ^9851      ^9861      ^9871      ^9881      ^9891      

ACCGCTTTTT TGCACAACAT GGGGGATCAT GTAACTCGCC TTGATCGTTG GGAACCGGAG CTGAATGAAG CCATACCAAA CGACGAGCGT GACACCACGA 
^9901      ^9911      ^9921      ^9931      ^9941      ^9951      ^9961      ^9971      ^9981      ^9991      

TGCCTGTAGC AATGGCAACA ACGTTGCGCA AACTATTAAC TGGCGAACTA CTTACTCTAG CTTCCCGGCA ACAATTAATA GACTGGATGG AGGCGGATAA 
^10001     ^10011     ^10021     ^10031     ^10041     ^10051     ^10061     ^10071     ^10081     ^10091     

AGTTGCAGGA CCACTTCTGC GCTCGGCCCT TCCGGCTGGC TGGTTTATTG CTGATAAATC TGGAGCCGGT GAGCGTGGGT CTCGCGGTAT CATTGCAGCA 
^10101     ^10111     ^10121     ^10131     ^10141     ^10151     ^10161     ^10171     ^10181     ^10191     

CTGGGGCCAG ATGGTAAGCC CTCCCGTATC GTAGTTATCT ACACGACGGG GAGTCAGGCA ACTATGGATG AACGAAATAG ACAGATCGCT GAGATAGGTG 
^10201     ^10211     ^10221     ^10231     ^10241     ^10251     ^10261     ^10271     ^10281     ^10291     

CCTCACTGAT TAAGCATTGG TAACTGTCAG ACCAAGTTTA CTCATATATA CTTTAGATTG ATTTAAAACT TCATTTTTAA TTTAAAAGGA TCTAGGTGAA 
^10301     ^10311     ^10321     ^10331     ^10341     ^10351     ^10361     ^10371     ^10381     ^10391     

GATCCTTTTT GATAATCTCA TGACCAAAAT CCCTTAACGT GAGTTTTCGT TCCACTGAGC GTCAGACCCC GTAGAAAAGA TCAAAGGATC TTCTTGAGAT 
^10401     ^10411     ^10421     ^10431     ^10441     ^10451     ^10461     ^10471     ^10481     ^10491     

CCTTTTTTTC TGCGCGTAAT CTGCTGCTTG CAAACAAAAA AACCACCGCT ACCAGCGGTG GTTTGTTTGC CGGATCAAGA GCTACCAACT CTTTTTCCGA 
^10501     ^10511     ^10521     ^10531     ^10541     ^10551     ^10561     ^10571     ^10581     ^10591     

AGGTAACTGG CTTCAGCAGA GCGCAGATAC CAAATACTGT TCTTCTAGTG TAGCCGTAGT TAGGCCACCA CTTCAAGAAC TCTGTAGCAC CGCCTACATA 
^10601     ^10611     ^10621     ^10631     ^10641     ^10651     ^10661     ^10671     ^10681     ^10691     

CCTCGCTCTG CTAATCCTGT TACCAGTGGC TGCTGCCAGT GGCGATAAGT CGTGTCTTAC CGGGTTGGAC TCAAGACGAT AGTTACCGGA TAAGGCGCAG 
^10701     ^10711     ^10721     ^10731     ^10741     ^10751     ^10761     ^10771     ^10781     ^10791     

CGGTCGGGCT GAACGGGGGG TTCGTGCACA CAGCCCAGCT TGGAGCGAAC GACCTACACC GAACTGAGAT ACCTACAGCG TGAGCTATGA GAAAGCGCCA 
^10801     ^10811     ^10821     ^10831     ^10841     ^10851     ^10861     ^10871     ^10881     ^10891     

CGCTTCCCGA AGGGAGAAAG GCGGACAGGT ATCCGGTAAG CGGCAGGGTC GGAACAGGAG AGCGCACGAG GGAGCTTCCA GGGGGAAACG CCTGGTATCT 
^10901     ^10911     ^10921     ^10931     ^10941     ^10951     ^10961     ^10971     ^10981     ^10991     

TTATAGTCCT GTCGGGTTTC GCCACCTCTG ACTTGAGCGT CGATTTTTGT GATGCTCGTC AGGGGGGCGG AGCCTATGGA AAAACGCCAG CAACGCGGCC 
^11001     ^11011     ^11021     ^11031     ^11041     ^11051     ^11061     ^11071     ^11081     ^11091     

TTTTTACGGT TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTTCTTTCC TGCGTTATCC CCTGATTCTG TGGATAACCG TATTACCGCC TTTGAGTGAG 
^11101     ^11111     ^11121     ^11131     ^11141     ^11151     ^11161     ^11171     ^11181     ^11191     

CTGATACCGC TCGCCGCAGC CGAACGACCG AGCGCAGCGA GTCAGTGAGC GAGGAAGCGG AAGAGCGCCC AATACGCAAA CCGCCTCTCC CCGCGCGTTG 
^11201     ^11211     ^11221     ^11231     ^11241     ^11251     ^11261     ^11271     ^11281     ^11291     

GCCGATTCAT TAATGCAGCT GGCACGACAG GTTTCCCGAC TGGAAAGCGG GCAGTGAGCG CAACGCAATT AATGTGAGTT AGCTCACTCA TTAGGCACCC 
^11301     ^11311     ^11321     ^11331     ^11341     ^11351     ^11361     ^11371     ^11381     ^11391     

CAGGCTTTAC ACTTTATGCT TCCGGCTCGT ATGTTGTGTG GAATTGTGAG CGGATAACAA TTTCACACAG GAAACAGCTA TGACCATGAT TACGCCAAGC 
^11401     ^11411     ^11421     ^11431     ^11441     ^11451     ^11461     ^11471     ^11481     ^11491     

TCGAAATTAA CCCTCACTAA AGGGAACAAA AGCTGGTACC GGGCCGGCCG TCGAGCTTGA TGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA 
^11501     ^11511     ^11521     ^11531     ^11541     ^11551     ^11561     ^11571     ^11581     ^11591     

TTCAGCTCCG GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC TCCGATCGTT GTCAGAAGTA 
^11601     ^11611     ^11621     ^11631     ^11641     ^11651     ^11661     ^11671     ^11681     ^11691     

AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC 
^11701     ^11711     ^11721     ^11731     ^11741     ^11751     ^11761     ^11771     ^11781     ^11791     

AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAC ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG 
^11801     ^11811     ^11821     ^11831     ^11841     ^11851     ^11861     ^11871     ^11881     ^11891     

CTCATCATTG GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC TCGTGCACCC AACTGATCTT 
^11901     ^11911     ^11921     ^11931     ^11941     ^11951     ^11961     ^11971     ^11981     ^11991     

CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT 
^12001     ^12011     ^12021     ^12031     ^12041     ^12051     ^12061     ^12071     ^12081     ^12091     

CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG 
^12101     ^12111     ^12121     ^12131     ^12141     ^12151     ^12161     ^12171     ^12181     ^12191     

GTTCCGCGCA CATTTCCCCG AAAAGTGCCA CCTGACGTCT AAGAAACCAT TATTATCATG ACATTAACCT ATAAAAATAG GCGTATCACG AGGCCCTTTC 
^12201     ^12211     ^12221     ^12231     ^12241     ^12251     ^12261     ^12271     ^12281     ^12291     

GTCTTCAAGA ATTGGGGATC TACGTATGGT CATTTCTTCT TCAGATTCCC TCATGGAGAA AGTGCGGCAG ATGTATATGA CAGAGTCGCC AGTTTCCAAG 
^12301     ^12311     ^12321     ^12331     ^12341     ^12351     ^12361     ^12371     ^12381     ^12391     

AGACTTTATT CAGGCACTTC CATGATAGGC AAGAGAGAAG ACCCAGAGAT GTTGTTGTCC TAGTTACACA TGGTATTTAT TCCAGAGTAT TCCTGATGAA 
^12401     ^12411     ^12421     ^12431     ^12441     ^12451     ^12461     ^12471     ^12481     ^12491     

ATGGTTTAGA TGGACATACG AAGAGTTTGA ATCGTTTACC AATGTTCCTA ACGGGAGCGT AATGGTGATG GAACTGGACG AATCCATCAA TAGATACGTC 
^12501     ^12511     ^12521     ^12531     ^12541     ^12551     ^12561     ^12571     ^12581     ^12591     

CTGAGGACCG TGCTACCCAA ATGGACTGAT TGTGAGGGAG ACCTAACTAC ATAGTGTTTA AAGATTACGG ATATTTAACT TACTTAGAAT AATGCCATTT 
^12601     ^12611     ^12621     ^12631     ^12641     ^12651     ^12661     ^12671     ^12681     ^12691     

TTTTGAGTTA TAATAATCCT ACGTTAGTGT GAGCGGGATT TAAACTGTGA GGACCTTAAT ACATTCAGAC ACTTCTGCGG TATCACCCTA CTTATTCCCT 
^12701     ^12711     ^12721     ^12731     ^12741     ^12751     ^12761     ^12771     ^12781     ^12791     

TCGAGATTAT ATCTAGGAAC CCATCAGGTT GGTGGAAGAT TACCCGTTCT AAGACTTTTC AGCTTCCTCT ATTGATGTTA CACCTGGACA CCCCTTTTCT 
^12801     ^12811     ^12821     ^12831     ^12841     ^12851     ^12861     ^12871     ^12881     ^12891     

GGCATCCAGT TTTTAATCTT CAGTGGCATG TGAGATTCTC CGAAATTAAT TAAAGCAATC ACACAATTCT CTCGGATACC ACCTCGGTTG AAACTGACAG 
^12901     ^12911     ^12921     ^12931     ^12941     ^12951     ^12961     ^12971     ^12981     ^12991     

GTGGTTTGTT ACGCATGCTA ATGCAAAGGA GCCTATATAC CTTTGGCTCG GCTGCTGTAA CAGGGAATAT AAAGGGCAGC ATAATTTAGG AGTTTAGTGA 
^13001     ^13011     ^13021     ^13031     ^13041     ^13051     ^13061     ^13071     ^13081     ^13091     

ACTTGCAACA TTTACTATTT TCCCTTCTTA CGTAAATATT TTTCTTTTTA ATTCTAAATC AATCTTTTTC AATTTTTTGT TTGTATTCTT TTCTTGCTTA 
^13101     ^13111     ^13121     ^13131     ^13141     ^13151     ^13161     ^13171     ^13181     ^13191     

AATCTATAAC TACAAAAAAC ACATACAG
^13201     ^13211     ^13221

Plasmid sequence verified (partially/completely) by IRESite (more details in Remarks):

plasmid sequence confirmed by IRESite curators by restriction analysis + parts by PCR + sequencing

GenBank formatted file with annotated plasmid sequence hyperlinked from vector image map:

The total number of notable open-reading frames (ORFs):

3

Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:

ORF

ORF position:

1

Version: `0`
Originaly submitted by: Tomáš Mašek	Reviewed by: Tomáš Mašek

The abbreviated name of this ORF/gene:
Fluc

The description of the protein encoded in this ORF:
firefly luciferase

The translational frameshift (ribosome slippage) involved:
0 The ribosome read-through involved:
no

The alternative forms of this protein occur by the alternative initiation of translation:

no

The ORF absolute position (the base range includes START and STOP codons or their equivalents):

1-1653

ORF

ORF position:

2

Version: `0`
Originaly submitted by: Tomáš Mašek	Reviewed by: Tomáš Mašek

The abbreviated name of this ORF/gene:
L270

The description of the protein encoded in this ORF:
arteficial short ORF derived from bacteriophage lambda DNA (25802-25746, GI:215104)

The translational frameshift (ribosome slippage) involved:
0 The ribosome read-through involved:
no

The alternative forms of this protein occur by the alternative initiation of translation:

no

The ORF absolute position (the base range includes START and STOP codons or their equivalents):

1744-1800

ORF

ORF position:

3

Version: `1`
Originaly submitted by: Tomáš Mašek	Reviewed by: Tomáš Mašek

The abbreviated name of this ORF/gene:
HCV1-GAL4

The description of the protein encoded in this ORF:
Fusion protein comprises the N-terminal 15 aminoacids of HCV polyprotein and the DNA-binding transcription factor Gal4, which is required for the activation of the GAL genes in response to galactose. Gal4 is repressed by Gal80p and activated by Gal3p.

The translational frameshift (ribosome slippage) involved:
0 The ribosome read-through involved:
no

The alternative forms of this protein occur by the alternative initiation of translation:

yes

The ORF absolute position (the base range includes START and STOP codons or their equivalents):

2261-4960

OPTIONAL: The function of the encoded protein

Molecular Function: transcriptional activator activity (TAS)
Biological Process: galactose metabolism (TAS) regulation of transcription, DNA-dependent (TAS)
Cellular Component: nucleus (TAS)

Remarks:

Sc_HCV_type_1a IRES was inserted into intercistronic position of pFGAL4-L270 tricistronic plasmid that can be
used in the pFGAL4h/PJ69A reporter system. The yeast strain PJ69A contains besides the GAL4 and GAL80
deletions also reporter genes coding for the yeast Ade2 and His3 proteins and for the bacterial
beta-galactosidase - all of them under the control of the Gal4-inducible Gal1 promoter. Thus pFGAL4/PJ69A
represents a specialised and sensitive system, which allows an enhancement of the measured signal by in vivo
coupled transcription and enzymatic detection. This system can be used for both measuring the ratio of the
beta-gal/luc enzymatic activities and searching for IRES activity by screening the colony growth rates on
selection media lacking adenine and histidine and containing various concentrations of the competitive
inhibitor of the histidine biosynthetic pathway.

Vector was created by IRESite curator - thus whole sequence is verified by above mentioned methods.

Citations:

Masek T., Vopalensky V., Horvath O., Vortelova L., Feketova Z., Pospisek M. (2007) Hepatitis C virus internal ribosome entry site initiates protein synthesis at the authentic initiation codon in yeast. J. Gen. Virol. 88(Pt 7):1992-2002

IRESs:

IRES:

Version: `3`	Last change: `2008-07-04 12:37:02`
Originaly submitted by: Tomáš Mašek	Reviewed by: Tomáš Mašek
The IRES name: `HCV_type_1a`	The functional status of IRES: `functional`

The IRES absolute position (the range includes START and STOP codons or their equivalents):

1920-2304

How IRES boundaries were determined:
experimentally_determined

5'-end of IRES relative to last base of the STOP codon of the upstream ORF: `120`	3'-end of IRES relative to last base of the STOP codon of the upstream ORF: `504`
5'-end of IRES relative to first base of the START codon of the downstream ORF: `-341`	3'-end of IRES relative to first base of the START codon of the downstream ORF: `43`

The sequence of IRES region aligned to its secondary structure (if available):

GCCAGCCCCC TGATGGGGGC GACACTCCAC CATGAATCAC TCCCCTGTGA GGAACTACTG TCTTCACGCA GAAAGCGTCT AGCCATGGCG TTAGTATGAG 
^1920      ^1930      ^1940      ^1950      ^1960      ^1970      ^1980      ^1990      ^2000      ^2010      

TGTCGTGCAG CCTCCAGGAC CCCCCCTCCC GGGAGAGCCA TAGTGGTCTG CGGAACCGGT GAGTACACCG GAATTGCCAG GACGACCGGG TCCTTTCTTG 
^2020      ^2030      ^2040      ^2050      ^2060      ^2070      ^2080      ^2090      ^2100      ^2110      

GATAAACCCG CTCAATGCCT GGAGATTTGG GCGTGCCCCC GCAAGACTGC TAGCCGAGTA GTGTTGGGTC GCGAAAGGCC TTGTGGTACT GCCTGATAGG 
^2120      ^2130      ^2140      ^2150      ^2160      ^2170      ^2180      ^2190      ^2200      ^2210      

GTGCTTGCGA GTGCCCCGGG AGGTCTCGTA GACCGTGCAC CATGAGCACG AATCCTAAAC CTCAAAGAAA AACCAAACGT AACAC
^2220      ^2230      ^2240      ^2250      ^2260      ^2270      ^2280      ^2290      ^2300

Remarks:

Sc_HCV_type_1a IRES represents the full-length 5'-UTR of HCV RNA (type 1a) and the first 15 codons of viral
polyprotein.

Citations:

The translation experiments:

Translation results:

IRESite Translation Id: 89

Version: `4`	Last change: `2008-07-04 12:37:02`
Originaly submitted by: Tomáš Mašek	Reviewed by: Tomáš Mašek

The translation method used to study IRES function:
in vivo

The organism used for translation:

Saccharomyces cerevisiae pJ69a

The temperature (in degrees of Celsia):
28

The relative translation efficiency in % of this IRES:

49.000

Name of IRES used as the positive control:

HCV1a

Name of the plasmid used as the positive control. `pFGAL4h-HCV1`	Name of the plasmid used as the negative control. `pFGAL4-L428`
`IRESite Id` of the plasmid used as positive control. `86`	`IRESite Id` of the plasmid used as negative control. `126`
The relative translation efficiency in % of the positive control: `100.000`	The relative translation efficiency in % of the negative control: `8.000`
The size (length) of intercistronic region in the positive control: `453`	The size (length) of intercistronic region in the negative control: `123`

The effect of 5'-cap analogs on translation:
not tested

Rapamycin affects translation:
not tested

Type of RNA subject to translation:

endogenous_nuclear_RNA_Pol_II_transcript

Remarks:

The relative translation efficiency and size of intercistronic region of negative control have been inserted
as an average values of all negative controls used (pFGAL4-L413,L-428, L456, IDs 126-128).

Citations:

Last change to the database: 2019-03-18 09:32:49 GMT+1