IRESite_Id:456 negative control plasmid pFGAL4h

IRESite: The database of experimentally verified IRES structures

The nucleic acid data:

IRESite Id: `456`	Version: `1`
Originaly submitted by: Tomáš Mašek
Reviewed by: Martin Mokrejš	Last change: `2008-09-12 14:51:30`

IRESite record type:

negative_control_plasmid_with_promoter_and_without_putative_IRES

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 bicistronic plasmid pFGAL4h which comprises luciferase and Gal4 genes as the first and
the second cistron, respectively. 13G:C hair pin loop was inserted between the two cistrons to prevent
ribosome read-through.

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 CTAGGAAACC CCCCCCCCCC CAAAGGGGGG GGGGGGGAGG 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

GCCCAGGATC CGTCGACAAG CTTGAATTCA AAAATGAAGC TACTGTCTTC TATCGAACAA GCATGCGATA TTTGCCGACT TAAAAAGCTC AAGTGCTCCA 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

AAGAAAAACC GAAGTGCGCC AAGTGTCTGA AGAACAACTG GGAGTGTCGC TACTCTCCCA AAACCAAAAG GTCTCCGCTG ACTAGGGCAC ATCTGACAGA 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

AGTGGAATCA AGGCTAGAAA GACTGGAACA GCTATTTCTA CTGATTTTTC CTCGAGAAGA CCTTGACATG ATTTTGAAAA TGGATTCTTT ACAGGATATA 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

AAAGCATTGT TAACAGGATT ATTTGTACAA GATAATGTGA ATAAAGATGC CGTCACAGAT AGATTGGCTT CAGTGGAGAC TGATATGCCT CTAACATTGA 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

GACAGCATAG AATAAGTGCG ACATCATCAT CGGAAGAGAG TAGTAACAAA GGTCAAAGAC AGTTGACTGT ATCGATTGAC TCGGCAGCTC ATCATGATAA 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

CTCCACAATT CCGTTGGATT TTATGCCCAG GGATGCTCTT CATGGATTTG ATTGGTCTGA AGAGGATGAC ATGTCGGATG GCTTGCCCTT CCTGAAAACG 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

GACCCCAACA ATAATGGGTT CTTTGGCGAC GGTTCTCTCT TATGTATTCT TCGATCTATT GGCTTTAAAC CGGAAAATTA CACGAACTCT AACGTTAACA 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

GGCTCCCGAC CATGATTACG GATAGATACA CGTTGGCTTC TAGATCCACA ACATCCCGTT TACTTCAAAG TTATCTCAAT AATTTTCACC CCTACTGCCC 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

TATCGTGCAC TCACCGACGC TAATGATGTT GTATAATAAC CAGATTGAAA TCGCGTCGAA GGATCAATGG CAAATCCTTT TTAACTGCAT ATTAGCCATT 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

GGAGCCTGGT GTATAGAGGG GGAATCTACT GATATAGATG TTTTTTACTA TCAAAATGCT AAATCTCATT TGACGAGCAA GGTCTTCGAG TCAGGTTCCA 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

TAATTTTGGT GACAGCCCTA CATCTTCTGT CGCGATATAC ACAGTGGAGG CAGAAAACAA ATACTAGCTA TAATTTTCAC AGCTTTTCCA TAAGAATGGC 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

CATATCATTG GGCTTGAATA GGGACCTCCC CTCGTCCTTC AGTGATAGCA GCATTCTGGA ACAAAGACGC CGAATTTGGT GGTCTGTCTA CTCTTGGGAG 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

ATCCAATTGT CCCTGCTTTA TGGTCGATCC ATCCAGCTTT CTCAGAATAC AATCTCCTTC CCTTCTTCTG TCGACGATGT GCAGCGTACC ACAACAGGTC 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

CCACCATATA TCATGGCATC ATTGAAACAG CAAGGCTCTT ACAAGTTTTC ACAAAAATCT ATGAACTAGA CAAAACAGTA ACTGCAGAAA AAAGTCCTAT 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

ATGTGCAAAA AAATGCTTGA TGATTTGTAA TGAGATTGAG GAGGTTTCGA GACAGGCACC AAAGTTTTTA CAAATGGATA TTTCCACCAC CGCTCTAACC 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

AATTTGTTGA AGGAACACCC TTGGCTATCC TTTACAAGAT TCGAACTGAA GTGGAAACAG TTGTCTCTTA TCATTTATGT ATTAAGAGAT TTTTTCACTA 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

ATTTTACCCA GAAAAAGTCA CAACTAGAAC AGGATCAAAA TGATCATCAA AGTTATGAAG TTAAACGATG CTCCATCATG TTAAGCGATG CAGCACAAAG 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

AACTGTTATG TCTGTAAGTA GCTATATGGA CAATCATAAT GTCACCCCAT ATTTTGCCTG GAATTGTTCT TATTACTTGT TCAATGCAGT CCTAGTACCC 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

ATAAAGACTC TACTCTCAAA CTCAAAATCG AATGCTGAGA ATAACGAGAC CGCACAATTA TTACAACAAA TTAACACTGT TCTGATGCTA TTAAAAAAAC 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

TGGCCACTTT TAAAATCCAG ACTTGTGAAA AATACATTCA AGTACTGGAA GAGGTATGTG CGCCGTTTCT GTTATCACAG TGTGCAATCC CATTACCGCA 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

TATCAGTTAT AACAATAGTA ATGGTAGCGC CATTAAAAAT ATTGTCGGTT CTGCAACTAT CGCCCAATAC CCTACTCTTC CGGAGGAAAA TGTCAACAAT 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

ATCAGTGTTA AATATGTTTC TCCTGGCTCA GTAGGGCCTT CACCTGTGCC ATTGAAATCA GGAGCAAGTT TCAGTGATCT AGTCAAGCTG TTATCTAACC 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

GTCCACCCTC TCGTAACTCT CCAGTGACAA TACCAAGAAG CACACCTTCG CATCGCTCAG TCACGCCTTT TCTAGGGCAA CAGCAACAGC TGCAATCATT 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

AGTGCCACTG ACCCCGTCTG CTTTGTTTGG TGGCGCCAAT TTTAATCAAA GTGGGAATAT TGCTGATAGC TCATTGTCCT TCACTTTCAC TAACAGTAGC 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

AACGGTCCGA ACCTCATAAC AACTCAAACA AATTCTCAAG CGCTTTCACA ACCAATTGCC TCCTCTAACG TTCATGATAA CTTCATGAAT AATGAAATCA 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

CGGCTAGTAA AATTGATGAT GGTAATAATT CAAAACCACT GTCACCTGGT TGGACGGACC AAACTGCGTA TAACGCGTTT GGAATCACTA CAGGGATGTT 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

TAATACCACT ACAATGGATG ATGTATATAA CTATCTATTC GATGATGAAG ATACCCCACC AAACCCAAAA AAAGAGTAA
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371

Credibility of mRNA sequence:

end-to-end_sequence_completely_same_as_in_the_experiment

The name of the plasmid:
pFGAL4h The name of the promoter used to express this mRNA:
TPI

Description of the plasmid (facultative for promoter-less plasmid records):
pFGAL4h represents yeast multi-copy expression shuttle vector and contains strong constitutive TPI promoter.

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

yes

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

no

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:
heterogeneous_population_of_molecules_found

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:
heterogeneous_population_of_molecules_found

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

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:
promoter_confirmed

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 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 AGCCTAGGAA ACCCCCCCCC CCCCAAAGGG 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

GGGGGGGGGG AGGGCCCAGG ATCCGTCGAC AAGCTTGAAT TCAAAAATGA AGCTACTGTC TTCTATCGAA CAAGCATGCG ATATTTGCCG ACTTAAAAAG 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

CTCAAGTGCT CCAAAGAAAA ACCGAAGTGC GCCAAGTGTC TGAAGAACAA CTGGGAGTGT CGCTACTCTC CCAAAACCAA AAGGTCTCCG CTGACTAGGG 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

CACATCTGAC AGAAGTGGAA TCAAGGCTAG AAAGACTGGA ACAGCTATTT CTACTGATTT TTCCTCGAGA AGACCTTGAC ATGATTTTGA AAATGGATTC 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

TTTACAGGAT ATAAAAGCAT TGTTAACAGG ATTATTTGTA CAAGATAATG TGAATAAAGA TGCCGTCACA GATAGATTGG CTTCAGTGGA GACTGATATG 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

CCTCTAACAT TGAGACAGCA TAGAATAAGT GCGACATCAT CATCGGAAGA GAGTAGTAAC AAAGGTCAAA GACAGTTGAC TGTATCGATT GACTCGGCAG 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

CTCATCATGA TAACTCCACA ATTCCGTTGG ATTTTATGCC CAGGGATGCT CTTCATGGAT TTGATTGGTC TGAAGAGGAT GACATGTCGG ATGGCTTGCC 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

CTTCCTGAAA ACGGACCCCA ACAATAATGG GTTCTTTGGC GACGGTTCTC TCTTATGTAT TCTTCGATCT ATTGGCTTTA AACCGGAAAA TTACACGAAC 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

TCTAACGTTA ACAGGCTCCC GACCATGATT ACGGATAGAT ACACGTTGGC TTCTAGATCC ACAACATCCC GTTTACTTCA AAGTTATCTC AATAATTTTC 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

ACCCCTACTG CCCTATCGTG CACTCACCGA CGCTAATGAT GTTGTATAAT AACCAGATTG AAATCGCGTC GAAGGATCAA TGGCAAATCC TTTTTAACTG 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

CATATTAGCC ATTGGAGCCT GGTGTATAGA GGGGGAATCT ACTGATATAG ATGTTTTTTA CTATCAAAAT GCTAAATCTC ATTTGACGAG CAAGGTCTTC 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

GAGTCAGGTT CCATAATTTT GGTGACAGCC CTACATCTTC TGTCGCGATA TACACAGTGG AGGCAGAAAA CAAATACTAG CTATAATTTT CACAGCTTTT 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

CCATAAGAAT GGCCATATCA TTGGGCTTGA ATAGGGACCT CCCCTCGTCC TTCAGTGATA GCAGCATTCT GGAACAAAGA CGCCGAATTT GGTGGTCTGT 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

CTACTCTTGG GAGATCCAAT TGTCCCTGCT TTATGGTCGA TCCATCCAGC TTTCTCAGAA TACAATCTCC TTCCCTTCTT CTGTCGACGA TGTGCAGCGT 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

ACCACAACAG GTCCCACCAT ATATCATGGC ATCATTGAAA CAGCAAGGCT CTTACAAGTT TTCACAAAAA TCTATGAACT AGACAAAACA GTAACTGCAG 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

AAAAAAGTCC TATATGTGCA AAAAAATGCT TGATGATTTG TAATGAGATT GAGGAGGTTT CGAGACAGGC ACCAAAGTTT TTACAAATGG ATATTTCCAC 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

CACCGCTCTA ACCAATTTGT TGAAGGAACA CCCTTGGCTA TCCTTTACAA GATTCGAACT GAAGTGGAAA CAGTTGTCTC TTATCATTTA TGTATTAAGA 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

GATTTTTTCA CTAATTTTAC CCAGAAAAAG TCACAACTAG AACAGGATCA AAATGATCAT CAAAGTTATG AAGTTAAACG ATGCTCCATC ATGTTAAGCG 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

ATGCAGCACA AAGAACTGTT ATGTCTGTAA GTAGCTATAT GGACAATCAT AATGTCACCC CATATTTTGC CTGGAATTGT TCTTATTACT TGTTCAATGC 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

AGTCCTAGTA CCCATAAAGA CTCTACTCTC AAACTCAAAA TCGAATGCTG AGAATAACGA GACCGCACAA TTATTACAAC AAATTAACAC TGTTCTGATG 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

CTATTAAAAA AACTGGCCAC TTTTAAAATC CAGACTTGTG AAAAATACAT TCAAGTACTG GAAGAGGTAT GTGCGCCGTT TCTGTTATCA CAGTGTGCAA 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

TCCCATTACC GCATATCAGT TATAACAATA GTAATGGTAG CGCCATTAAA AATATTGTCG GTTCTGCAAC TATCGCCCAA TACCCTACTC TTCCGGAGGA 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

AAATGTCAAC AATATCAGTG TTAAATATGT TTCTCCTGGC TCAGTAGGGC CTTCACCTGT GCCATTGAAA TCAGGAGCAA GTTTCAGTGA TCTAGTCAAG 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

CTGTTATCTA ACCGTCCACC CTCTCGTAAC TCTCCAGTGA CAATACCAAG AAGCACACCT TCGCATCGCT CAGTCACGCC TTTTCTAGGG CAACAGCAAC 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

AGCTGCAATC ATTAGTGCCA CTGACCCCGT CTGCTTTGTT TGGTGGCGCC AATTTTAATC AAAGTGGGAA TATTGCTGAT AGCTCATTGT CCTTCACTTT 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

CACTAACAGT AGCAACGGTC CGAACCTCAT AACAACTCAA ACAAATTCTC AAGCGCTTTC ACAACCAATT GCCTCCTCTA ACGTTCATGA TAACTTCATG 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

AATAATGAAA TCACGGCTAG TAAAATTGAT GATGGTAATA ATTCAAAACC ACTGTCACCT GGTTGGACGG ACCAAACTGC GTATAACGCG TTTGGAATCA 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

CTACAGGGAT GTTTAATACC ACTACAATGG ATGATGTATA TAACTATCTA TTCGATGATG AAGATACCCC ACCAAACCCA AAAAAAGAGT AAGAGCTCAG 
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371      ^4381      ^4391      

CTAGCTAACT GAATAAGGAA CAATGAACGT TTTTCCTTTC TCTTGTTCCT AGTATTAATG ACTGACCGAT ACATCCCTTT TTTTTTTTGT CTTTGTCTAG 
^4401      ^4411      ^4421      ^4431      ^4441      ^4451      ^4461      ^4471      ^4481      ^4491      

CTCCAATTCG CCCTATAGTG AGTCGTATTA CAATTCACTG GCCGTCGTTT TACAACGTCG TGACTGGGAA AACCCTGGCG TTACCCAACT TAATCGCCTT 
^4501      ^4511      ^4521      ^4531      ^4541      ^4551      ^4561      ^4571      ^4581      ^4591      

GCAGCACATC CCCCTTTCGC CAGCTGGCGT AATAGCGAAG AGGCCCGCAC CGATCGCCCT TCCCAACAGT TGCGCAGCCT GAATGGCGAA TGGACGCGCC 
^4601      ^4611      ^4621      ^4631      ^4641      ^4651      ^4661      ^4671      ^4681      ^4691      

CTGTAGCGGC GCATTAAGCG CGGCGGGTGT GGTGGTTACG CGCAGCGTGA CCGCTACACT TGCCAGCGCC CTAGCGCCCG CTCCTTTCGC TTTCTTCCCT 
^4701      ^4711      ^4721      ^4731      ^4741      ^4751      ^4761      ^4771      ^4781      ^4791      

TCCTTTCTCG CCACGTTCGC CGGCTTTCCC CGTCAAGCTC TAAATCGGGG GCTCCCTTTA GGGTTCCGAT TTAGTGCTTT ACGGCACCTC GACCCCAAAA 
^4801      ^4811      ^4821      ^4831      ^4841      ^4851      ^4861      ^4871      ^4881      ^4891      

AACTTGATTA GGGTGATGGT TCACGTAGTG GGCCATCGCC CTGATAGACG GTTTTTCGCC CTTTGACGTT GGAGTCCACG TTCTTTAATA GTGGACTCTT 
^4901      ^4911      ^4921      ^4931      ^4941      ^4951      ^4961      ^4971      ^4981      ^4991      

GTTCCAAACT GGAACAACAC TCAACCCTAT CTCGGTCTAT TCTTTTGATT TATAAGGGAT TTTGCCGATT TCGGCCTATT GGTTAAAAAA TGAGCTGATT 
^5001      ^5011      ^5021      ^5031      ^5041      ^5051      ^5061      ^5071      ^5081      ^5091      

TAACAAAAAT TTAACGCGAA TTTTAACAAA ATATTAACGC TTACAATTTC CTGATGCGGT ATTTTCTCCT TACGCATCTG TGCGGTATTT CACACCGCAT 
^5101      ^5111      ^5121      ^5131      ^5141      ^5151      ^5161      ^5171      ^5181      ^5191      

AGGGTAATAA CTGATATAAT TAAATTGAAG CTCTAATTTG TGAGTTTAGT ATACATGCAT TTACTTATAA TACAGTTTTT TAGTTTTGCT GGCCGCATCT 
^5201      ^5211      ^5221      ^5231      ^5241      ^5251      ^5261      ^5271      ^5281      ^5291      

TCTCAAATAT GCTTCCCAGC CTGCTTTTCT GTAACGTTCA CCCTCTACCT TAGCATCCCT TCCCTTTGCA AATAGTCCTC TTCCAACAAT AATAATGTCA 
^5301      ^5311      ^5321      ^5331      ^5341      ^5351      ^5361      ^5371      ^5381      ^5391      

GATCCTGTAG AGACCACATC ATCCACGGTT CTATACTGTT GACCCAATGC GTCTCCCTTG TCATCTAAAC CCACACCGGG TGTCATAATC AACCAATCGT 
^5401      ^5411      ^5421      ^5431      ^5441      ^5451      ^5461      ^5471      ^5481      ^5491      

AACCTTCATC TCTTCCACCC ATGTCTCTTT GAGCAATAAA GCCGATAACA AAATCTTTGT CGCTCTTCGC AATGTCAACA GTACCCTTAG TATATTCTCC 
^5501      ^5511      ^5521      ^5531      ^5541      ^5551      ^5561      ^5571      ^5581      ^5591      

AGTAGATAGG GAGCCCTTGC ATGACAATTC TGCTAACATC AAAAGGCCTC TAGGTTCCTT TGTTACTTCT TCTGCCGCCT GCTTCAAACC GCTAACAATA 
^5601      ^5611      ^5621      ^5631      ^5641      ^5651      ^5661      ^5671      ^5681      ^5691      

CCTGGCCCCA CCACACCGTG TGCATTCGTA ATGTCTGCCC ATTCTGCTAT TCTGTATACA CCCGCAGAGT ACTGCAATTT GACTGTATTA CCAATGTCAG 
^5701      ^5711      ^5721      ^5731      ^5741      ^5751      ^5761      ^5771      ^5781      ^5791      

CAAATTTTCT GTCTTCGAAG AGTAAAAAAT TGTACTTGGC GGATAATGCC TTTAGCGGCT TAACTGTGCC CTCCATCGAA AAATCAGTCA ATATATCCAC 
^5801      ^5811      ^5821      ^5831      ^5841      ^5851      ^5861      ^5871      ^5881      ^5891      

ATGTGTTTTT AGTAAACAAA TTTTGGGACC TAATGCTTCA ACTAACTCCA GTAATTCCTT GGTGGTACGA ACATCCAATG AAGCACACAA GTTTGTTTGC 
^5901      ^5911      ^5921      ^5931      ^5941      ^5951      ^5961      ^5971      ^5981      ^5991      

TTTTCGTGCA TGATATTAAA TAGCTTGGCA GCAACAGGAC TAGGATGAGT AGCAGCACGT TCCTTATATG TAGCTTTCGA CATGATTTAT CTTCGTTTCC 
^6001      ^6011      ^6021      ^6031      ^6041      ^6051      ^6061      ^6071      ^6081      ^6091      

TGCAGGTTTT TGTTCTGTGC AGTTGGGTTA AGAATACTGG GCAATTTCAT GTTTCTTCAA CACTACATAT GCGTATATAT ACCAATCTAA GTCTGTGCTC 
^6101      ^6111      ^6121      ^6131      ^6141      ^6151      ^6161      ^6171      ^6181      ^6191      

CTTCCTTCGT TCTTCCTTCT GTTCGGAGAT TACCGAATCA AAAAAATTTC AAGGAAACCG AAATCAAAAA AAAGAATAAA AAAAAAATGA TGAATTGAAA 
^6201      ^6211      ^6221      ^6231      ^6241      ^6251      ^6261      ^6271      ^6281      ^6291      

AGGTGGTATG GTGCACTCTC AGTACAATCT GCTCTGATGC CGCATAGTTA AGCCAGCCCC GACACCCGCC AACACCCGCT GACGCGCCCT GACGGGCTTG 
^6301      ^6311      ^6321      ^6331      ^6341      ^6351      ^6361      ^6371      ^6381      ^6391      

TCTGCTCCCG GCATCCGCTT ACAGACAAGC TGTGACCGTC TCCGGGAGCT GCATGTGTCA GAGGTTTTCA CCGTCATCAC CGAAACGCGC GAGACGAAAG 
^6401      ^6411      ^6421      ^6431      ^6441      ^6451      ^6461      ^6471      ^6481      ^6491      

GGCCTCGTGA TACGCCTATT TTTATAGGTT AATGTCATGA TAATAATGGT TTCTTAGACG TGCGGCCGCT CTAGAACTAG TGGATCAATT CCACGGACTA 
^6501      ^6511      ^6521      ^6531      ^6541      ^6551      ^6561      ^6571      ^6581      ^6591      

TAGACTATAC TAGTATACTC CGTCTACTGT ACGATACACT TCCGCTCAGG TCCTTGTCCT TTAACGAGGC CTTACCACTC TTTTGTTACT CTATTGATCC 
^6601      ^6611      ^6621      ^6631      ^6641      ^6651      ^6661      ^6671      ^6681      ^6691      

AGCTCAGCAA AGGCAGTGTG ATCTAAGATT CTATCTTCGC GATGTAGTAA AACTAGCTAG ACCGAGAAAG AGACTAGAAA TGCAAAAGGC ACTTCTACAA 
^6701      ^6711      ^6721      ^6731      ^6741      ^6751      ^6761      ^6771      ^6781      ^6791      

TGGCTGCCAT CATTATTATC CGATGTGACG CTGCAGCTTC TCAATGATAT TCGAATACGC TTTGAGGAGA TACAGCCTAA TATCCGACAA ACTGTTTTAC 
^6801      ^6811      ^6821      ^6831      ^6841      ^6851      ^6861      ^6871      ^6881      ^6891      

AGATTTACGA TCGTACTTGT TACCCATCAT TGAATTTTGA ACATCCGAAC CTGGGAGTTT TCCCTGAAAC AGATAGTATA TTTGAACCTG TATAATAATA 
^6901      ^6911      ^6921      ^6931      ^6941      ^6951      ^6961      ^6971      ^6981      ^6991      

TATAGTCTAG CGCTTTACGG AAGACAATGT ATGTATTTCG GTTCCTGGAG AAACTATTGC ATCTATTGCA TAGGTAATCT TGCACGTCGC ATCCCCGGTT 
^7001      ^7011      ^7021      ^7031      ^7041      ^7051      ^7061      ^7071      ^7081      ^7091      

CATTTTCTGC GTTTCCATCT TGCACTTCAA TAGCATATCT TTGTTAACGA AGCATCTGTG CTTCATTTTG TAGAACAAAA ATGCAACGCG AGAGCGCTAA 
^7101      ^7111      ^7121      ^7131      ^7141      ^7151      ^7161      ^7171      ^7181      ^7191      

TTTTTCAAAC AAAGAATCTG AGCTGCATTT TTACAGAACA GAAATGCAAC GCGAAAGCGC TATTTTACCA ACGAAGAATC TGTGCTTCAT TTTTGTAAAA 
^7201      ^7211      ^7221      ^7231      ^7241      ^7251      ^7261      ^7271      ^7281      ^7291      

CAAAAATGCA ACGCGAGAGC GCTAATTTTT CAAACAAAGA ATCTGAGCTG CATTTTTACA GAACAGAAAT GCAACGCGAG AGCGCTATTT TACCAACAAA 
^7301      ^7311      ^7321      ^7331      ^7341      ^7351      ^7361      ^7371      ^7381      ^7391      

GAATCTATAC TTCTTTTTTG TTCTACAAAA ATGCATCCCG AGAGCGCTAT TTTTCTAACA AAGCATCTTA GATTACTTTT TTTCTCCTTT GTGCGCTCTA 
^7401      ^7411      ^7421      ^7431      ^7441      ^7451      ^7461      ^7471      ^7481      ^7491      

TAATGCAGTC TCTTGATAAC TTTTTGCACT GTAGGTCCGT TAAGGTTAGA AGAAGGCTAC TTTGGTGTCT ATTTTCTCTT CCATAAAAAA AGCCTGACTC 
^7501      ^7511      ^7521      ^7531      ^7541      ^7551      ^7561      ^7571      ^7581      ^7591      

CACTTCCCGC GTTTACTGAT TACTAGCGAA GCTGCGGGTG CATTTTTTCA AGATAAAGGC ATCCCCGATT ATATTCTATA CCGATGTGGA TTGCGCATAC 
^7601      ^7611      ^7621      ^7631      ^7641      ^7651      ^7661      ^7671      ^7681      ^7691      

TTTGTGAACA GAAAGTGATA GCGTTGATGA TTCTTCATTG GTCAGAAAAT TATGAACGGT TTCTTCTATT TTGTCTCTAT ATACTACGTA TAGGAAATGT 
^7701      ^7711      ^7721      ^7731      ^7741      ^7751      ^7761      ^7771      ^7781      ^7791      

TTACATTTTC GTATTGTTTT CGATTCACTC TATGAATAGT TCTTACTACA ATTTTTTTGT CTAAAGAGTA ATACTAGAGA TAAACATAAA AAATGTAGAG 
^7801      ^7811      ^7821      ^7831      ^7841      ^7851      ^7861      ^7871      ^7881      ^7891      

GTCGAGTTTA GATGCAAGTT CAAGGAGCGA AAGGTGGATG GGTAGGTTAT ATAGGGATAT AGCACAGAGA TATATAGCAA AGAGATACTT TTGAGCAATG 
^7901      ^7911      ^7921      ^7931      ^7941      ^7951      ^7961      ^7971      ^7981      ^7991      

TTTGTGGAAG CGGTATTCGC AATATTTTAG TAGCTCGTTA CAGTCCGGTG CGTTTTTGGT TTTTTGAAAG TGCGTCTTCA GAGCGCTTTT GGTTTTCAAA 
^8001      ^8011      ^8021      ^8031      ^8041      ^8051      ^8061      ^8071      ^8081      ^8091      

AGCGCTCTGA AGTTCCTATA CTTTCTAGAG AATAGGAACT TCGGAATAGG AACTTCAAAG CGTTTCCGAA AACGAGCGCT TCCGAAAATG CAACGCGAGC 
^8101      ^8111      ^8121      ^8131      ^8141      ^8151      ^8161      ^8171      ^8181      ^8191      

TGCGCACATA CAGCTCACTG TTCACGTCGC ACCTATATCT GCGTGTTGCC TGTATATATA TATACATGAG AAGAACGGCA TAGTGCGTGT TTATGCTTAA 
^8201      ^8211      ^8221      ^8231      ^8241      ^8251      ^8261      ^8271      ^8281      ^8291      

ATGCGTACTT ATATGCGTCT ATTTATGTAG GATGAAAGGT AGTCTAGTAC CTCCTGTGAT ATTATCCCAT TCCATGCGGG GTATCGTATG CTTCCTTCAG 
^8301      ^8311      ^8321      ^8331      ^8341      ^8351      ^8361      ^8371      ^8381      ^8391      

CACTACCCTT TAGCTGTTCT ATATGCTGCC ACTCCTCAAT TGGATTAGTC TCATCCTTCA ATGCTATCAT TTCCTTTGAT ATTGGATCAT ATGCATAGTA 
^8401      ^8411      ^8421      ^8431      ^8441      ^8451      ^8461      ^8471      ^8481      ^8491      

CCGAGAAACT AGTGCGAAGT AGTGATCAGG TATTGCTGTT ATCTGATGAG TATACGTTGT CCTGGCCACG GCAGAAGCAC GCTTATCGCT CCAATTTCCC 
^8501      ^8511      ^8521      ^8531      ^8541      ^8551      ^8561      ^8571      ^8581      ^8591      

ACAACATTAG TCAACTCCGT TAGGCCCTTC ATTGAAAGAA ATGAGGTCAT CAAATGTCTT CCAATGTGAG ATTTTGGGCC ATTTTTTATA GCAAAGATTG 
^8601      ^8611      ^8621      ^8631      ^8641      ^8651      ^8661      ^8671      ^8681      ^8691      

AATAAGGCGC ATTTTTCTTC AAAGCTGCGG CCGCACTCTC ACTAGTACGT CAGGTGGCAC TTTTCGGGGA AATGTGCGCG GAACCCCTAT TTGTTTATTT 
^8701      ^8711      ^8721      ^8731      ^8741      ^8751      ^8761      ^8771      ^8781      ^8791      

TTCTAAATAC ATTCAAATAT GTATCCGCTC ATGAGACAAT AACCCTGATA AATGCTTCAA TAATATTGAA AAAGGAAGAG TATGAGTATT CAACATTTCC 
^8801      ^8811      ^8821      ^8831      ^8841      ^8851      ^8861      ^8871      ^8881      ^8891      

GTGTCGCCCT TATTCCCTTT TTTGCGGCAT TTTGCCTTCC TGTTTTTGCT CACCCAGAAA CGCTGGTGAA AGTAAAAGAT GCTGAAGATC AGTTGGGTGC 
^8901      ^8911      ^8921      ^8931      ^8941      ^8951      ^8961      ^8971      ^8981      ^8991      

ACGAGTGGGT TACATCGAAC TGGATCTCAA CAGCGGTAAG ATCCTTGAGA GTTTTCGCCC CGAAGAACGT TTTCCAATGA TGAGCACTTT TAAAGTTCTG 
^9001      ^9011      ^9021      ^9031      ^9041      ^9051      ^9061      ^9071      ^9081      ^9091      

CTATGTGGCG CGGTATTATC CCGTATTGAC GCCGGGCAAG AGCAACTCGG TCGCCGCATA CACTATTCTC AGAATGACTT GGTTGAGTAC TCACCAGTCA 
^9101      ^9111      ^9121      ^9131      ^9141      ^9151      ^9161      ^9171      ^9181      ^9191      

CAGAAAAGCA TCTTACGGAT GGCATGACAG TAAGAGAATT ATGCAGTGCT GCCATAACCA TGAGTGATAA CACTGCGGCC AACTTACTTC TGACAACGAT 
^9201      ^9211      ^9221      ^9231      ^9241      ^9251      ^9261      ^9271      ^9281      ^9291      

CGGAGGACCG AAGGAGCTAA CCGCTTTTTT GCACAACATG GGGGATCATG TAACTCGCCT TGATCGTTGG GAACCGGAGC TGAATGAAGC CATACCAAAC 
^9301      ^9311      ^9321      ^9331      ^9341      ^9351      ^9361      ^9371      ^9381      ^9391      

GACGAGCGTG ACACCACGAT GCCTGTAGCA ATGGCAACAA CGTTGCGCAA ACTATTAACT GGCGAACTAC TTACTCTAGC TTCCCGGCAA CAATTAATAG 
^9401      ^9411      ^9421      ^9431      ^9441      ^9451      ^9461      ^9471      ^9481      ^9491      

ACTGGATGGA GGCGGATAAA GTTGCAGGAC CACTTCTGCG CTCGGCCCTT CCGGCTGGCT GGTTTATTGC TGATAAATCT GGAGCCGGTG AGCGTGGGTC 
^9501      ^9511      ^9521      ^9531      ^9541      ^9551      ^9561      ^9571      ^9581      ^9591      

TCGCGGTATC ATTGCAGCAC TGGGGCCAGA TGGTAAGCCC TCCCGTATCG TAGTTATCTA CACGACGGGG AGTCAGGCAA CTATGGATGA ACGAAATAGA 
^9601      ^9611      ^9621      ^9631      ^9641      ^9651      ^9661      ^9671      ^9681      ^9691      

CAGATCGCTG AGATAGGTGC CTCACTGATT AAGCATTGGT AACTGTCAGA CCAAGTTTAC TCATATATAC TTTAGATTGA TTTAAAACTT CATTTTTAAT 
^9701      ^9711      ^9721      ^9731      ^9741      ^9751      ^9761      ^9771      ^9781      ^9791      

TTAAAAGGAT CTAGGTGAAG ATCCTTTTTG ATAATCTCAT GACCAAAATC CCTTAACGTG AGTTTTCGTT CCACTGAGCG TCAGACCCCG TAGAAAAGAT 
^9801      ^9811      ^9821      ^9831      ^9841      ^9851      ^9861      ^9871      ^9881      ^9891      

CAAAGGATCT TCTTGAGATC CTTTTTTTCT GCGCGTAATC TGCTGCTTGC AAACAAAAAA ACCACCGCTA CCAGCGGTGG TTTGTTTGCC GGATCAAGAG 
^9901      ^9911      ^9921      ^9931      ^9941      ^9951      ^9961      ^9971      ^9981      ^9991      

CTACCAACTC TTTTTCCGAA GGTAACTGGC TTCAGCAGAG CGCAGATACC AAATACTGTT CTTCTAGTGT AGCCGTAGTT AGGCCACCAC TTCAAGAACT 
^10001     ^10011     ^10021     ^10031     ^10041     ^10051     ^10061     ^10071     ^10081     ^10091     

CTGTAGCACC GCCTACATAC CTCGCTCTGC TAATCCTGTT ACCAGTGGCT GCTGCCAGTG GCGATAAGTC GTGTCTTACC GGGTTGGACT CAAGACGATA 
^10101     ^10111     ^10121     ^10131     ^10141     ^10151     ^10161     ^10171     ^10181     ^10191     

GTTACCGGAT AAGGCGCAGC GGTCGGGCTG AACGGGGGGT TCGTGCACAC AGCCCAGCTT GGAGCGAACG ACCTACACCG AACTGAGATA CCTACAGCGT 
^10201     ^10211     ^10221     ^10231     ^10241     ^10251     ^10261     ^10271     ^10281     ^10291     

GAGCTATGAG AAAGCGCCAC GCTTCCCGAA GGGAGAAAGG CGGACAGGTA TCCGGTAAGC GGCAGGGTCG GAACAGGAGA GCGCACGAGG GAGCTTCCAG 
^10301     ^10311     ^10321     ^10331     ^10341     ^10351     ^10361     ^10371     ^10381     ^10391     

GGGGAAACGC CTGGTATCTT TATAGTCCTG TCGGGTTTCG CCACCTCTGA CTTGAGCGTC GATTTTTGTG ATGCTCGTCA GGGGGGCGGA GCCTATGGAA 
^10401     ^10411     ^10421     ^10431     ^10441     ^10451     ^10461     ^10471     ^10481     ^10491     

AAACGCCAGC AACGCGGCCT TTTTACGGTT CCTGGCCTTT TGCTGGCCTT TTGCTCACAT GTTCTTTCCT GCGTTATCCC CTGATTCTGT GGATAACCGT 
^10501     ^10511     ^10521     ^10531     ^10541     ^10551     ^10561     ^10571     ^10581     ^10591     

ATTACCGCCT TTGAGTGAGC TGATACCGCT CGCCGCAGCC GAACGACCGA GCGCAGCGAG TCAGTGAGCG AGGAAGCGGA AGAGCGCCCA ATACGCAAAC 
^10601     ^10611     ^10621     ^10631     ^10641     ^10651     ^10661     ^10671     ^10681     ^10691     

CGCCTCTCCC CGCGCGTTGG CCGATTCATT AATGCAGCTG GCACGACAGG TTTCCCGACT GGAAAGCGGG CAGTGAGCGC AACGCAATTA ATGTGAGTTA 
^10701     ^10711     ^10721     ^10731     ^10741     ^10751     ^10761     ^10771     ^10781     ^10791     

GCTCACTCAT TAGGCACCCC AGGCTTTACA CTTTATGCTT CCGGCTCGTA TGTTGTGTGG AATTGTGAGC GGATAACAAT TTCACACAGG AAACAGCTAT 
^10801     ^10811     ^10821     ^10831     ^10841     ^10851     ^10861     ^10871     ^10881     ^10891     

GACCATGATT ACGCCAAGCT CGAAATTAAC CCTCACTAAA GGGAACAAAA GCTGGTACCG GGCCGGCCGT CGAGCTTGAT GGCATCGTGG TGTCACGCTC 
^10901     ^10911     ^10921     ^10931     ^10941     ^10951     ^10961     ^10971     ^10981     ^10991     

GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT 
^11001     ^11011     ^11021     ^11031     ^11041     ^11051     ^11061     ^11071     ^11081     ^11091     

CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTG CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT 
^11101     ^11111     ^11121     ^11131     ^11141     ^11151     ^11161     ^11171     ^11181     ^11191     

CTGTGACTGG TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC GGCGTCAACA CGGGATAATA CCGCGCCACA 
^11201     ^11211     ^11221     ^11231     ^11241     ^11251     ^11261     ^11271     ^11281     ^11291     

TAGCAGAACT TTAAAAGTGC TCATCATTGG AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACT 
^11301     ^11311     ^11321     ^11331     ^11341     ^11351     ^11361     ^11371     ^11381     ^11391     

CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAA ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA 
^11401     ^11411     ^11421     ^11431     ^11441     ^11451     ^11461     ^11471     ^11481     ^11491     

CACGGAAATG TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT CATGAGCGGA TACATATTTG AATGTATTTA 
^11501     ^11511     ^11521     ^11531     ^11541     ^11551     ^11561     ^11571     ^11581     ^11591     

GAAAAATAAA CAAATAGGGG TTCCGCGCAC ATTTCCCCGA AAAGTGCCAC CTGACGTCTA AGAAACCATT ATTATCATGA CATTAACCTA TAAAAATAGG 
^11601     ^11611     ^11621     ^11631     ^11641     ^11651     ^11661     ^11671     ^11681     ^11691     

CGTATCACGA GGCCCTTTCG TCTTCAAGAA TTGGGGATCT ACGTATGGTC ATTTCTTCTT CAGATTCCCT CATGGAGAAA GTGCGGCAGA TGTATATGAC 
^11701     ^11711     ^11721     ^11731     ^11741     ^11751     ^11761     ^11771     ^11781     ^11791     

AGAGTCGCCA GTTTCCAAGA GACTTTATTC AGGCACTTCC ATGATAGGCA AGAGAGAAGA CCCAGAGATG TTGTTGTCCT AGTTACACAT GGTATTTATT 
^11801     ^11811     ^11821     ^11831     ^11841     ^11851     ^11861     ^11871     ^11881     ^11891     

CCAGAGTATT CCTGATGAAA TGGTTTAGAT GGACATACGA AGAGTTTGAA TCGTTTACCA ATGTTCCTAA CGGGAGCGTA ATGGTGATGG AACTGGACGA 
^11901     ^11911     ^11921     ^11931     ^11941     ^11951     ^11961     ^11971     ^11981     ^11991     

ATCCATCAAT AGATACGTCC TGAGGACCGT GCTACCCAAA TGGACTGATT GTGAGGGAGA CCTAACTACA TAGTGTTTAA AGATTACGGA TATTTAACTT 
^12001     ^12011     ^12021     ^12031     ^12041     ^12051     ^12061     ^12071     ^12081     ^12091     

ACTTAGAATA ATGCCATTTT TTTGAGTTAT AATAATCCTA CGTTAGTGTG AGCGGGATTT AAACTGTGAG GACCTTAATA CATTCAGACA CTTCTGCGGT 
^12101     ^12111     ^12121     ^12131     ^12141     ^12151     ^12161     ^12171     ^12181     ^12191     

ATCACCCTAC TTATTCCCTT CGAGATTATA TCTAGGAACC CATCAGGTTG GTGGAAGATT ACCCGTTCTA AGACTTTTCA GCTTCCTCTA TTGATGTTAC 
^12201     ^12211     ^12221     ^12231     ^12241     ^12251     ^12261     ^12271     ^12281     ^12291     

ACCTGGACAC CCCTTTTCTG GCATCCAGTT TTTAATCTTC AGTGGCATGT GAGATTCTCC GAAATTAATT AAAGCAATCA CACAATTCTC TCGGATACCA 
^12301     ^12311     ^12321     ^12331     ^12341     ^12351     ^12361     ^12371     ^12381     ^12391     

CCTCGGTTGA AACTGACAGG TGGTTTGTTA CGCATGCTAA TGCAAAGGAG CCTATATACC TTTGGCTCGG CTGCTGTAAC AGGGAATATA AAGGGCAGCA 
^12401     ^12411     ^12421     ^12431     ^12441     ^12451     ^12461     ^12471     ^12481     ^12491     

TAATTTAGGA GTTTAGTGAA CTTGCAACAT TTACTATTTT CCCTTCTTAC GTAAATATTT TTCTTTTTAA TTCTAAATCA ATCTTTTTCA ATTTTTTGTT 
^12501     ^12511     ^12521     ^12531     ^12541     ^12551     ^12561     ^12571     ^12581     ^12591     

TGTATTCTTT TCTTGCTTAA ATCTATAACT ACAAAAAACA CATACAG
^12601     ^12611     ^12621     ^12631     ^12641

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

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

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

A promoter reported in cDNA corresponding to IRES sequence:
not tested The total number of notable open-reading frames (ORFs):
2

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: Martin Mokrejš

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: Martin Mokrejš

The abbreviated name of this ORF/gene:
Gal4

The description of the protein encoded in this ORF:
GAL4 is a DNA-binding transcription factor. Gal4p is required for the activation of the GAL genes in response to galactose; 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:

no

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

1734-4379

Remarks:

pFGAL4h bicistronic plasmid has been designed for the use in the pFGAL4h/PJ69A reporter system. pFGAL4h vector
bears genes for the firefly luciferase and Gal4p as a first and second cistron, respectively.
A stable stem-loop hairpin containing 13G:C pairs was inserted between the two genes in order to prevent
ribosome read-through from the first to the second cistron. 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.

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

Vopalensky V., Masek T., Horvath O., Vicenova B., Mokrejs M., Pospisek M. (2008) Firefly luciferase gene contains a cryptic promoter. RNA. 14(9):1720-1729

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