IRESite_Id:530 characteristics of c-IAP1_-150/-1 IRES studied in pbetaGAL/HIAP2(1073-1222)/CAT plasmid

IRESite: The database of experimentally verified IRES structures

The nucleic acid data:

IRESite Id: `530`	Version: `5`
Originaly submitted by: Václav Vopálenský
Reviewed by: Martin Mokrejš	Last change: `2008-10-27 23:35:24`

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:
3UTR_possibly_incomplete

The mRNA/+RNA description:

Putative in vivo CMV promoter-derived transcript produced from bicistronic plasmid
pbetaGAL/HIAP2(1073-1222)/CAT which comprises beta galactosidase and chloramphenicol acetyltransferase as the
first and the second cistron respectively and the part of human c-IAP1 5' UTR (nt from -150 to nt -1 of the
original sequence) mRNA mRNA cloned between them.
The sequence ends at its 3'-end right after the poly(A) signal from BGH mRNA and thus the 3'-UTR might be
slightly wrong.

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

agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ccactagtaa cggccgccag 
^1         ^11        ^21        ^31        ^41        ^51        ^61        ^71        ^81        ^91        

tgtgctggaa ttctgcagat atccatcaca ctggcggccg caattcccgg ggatcgaaag agcctgctaa agcaaaaaag aagtcaccat gtcgtttact 
^101       ^111       ^121       ^131       ^141       ^151       ^161       ^171       ^181       ^191       

ttgaccaaca agaacgtgat tttcgttgcc ggtctgggag gcattggtct ggacaccagc aaggagctgc tcaagcgcga tcccgtcgtt ttacaacgtc 
^201       ^211       ^221       ^231       ^241       ^251       ^261       ^271       ^281       ^291       

gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc 
^301       ^311       ^321       ^331       ^341       ^351       ^361       ^371       ^381       ^391       

ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag 
^401       ^411       ^421       ^431       ^441       ^451       ^461       ^471       ^481       ^491       

gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa cctatcccat tacggtcaat ccgccgtttg 
^501       ^511       ^521       ^531       ^541       ^551       ^561       ^571       ^581       ^591       

ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttaa 
^601       ^611       ^621       ^631       ^641       ^651       ^661       ^671       ^681       ^691       

ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg acagtcgttt gccgtctgaa tttgacctga gcgcattttt acgcgccgga 
^701       ^711       ^721       ^731       ^741       ^751       ^761       ^771       ^781       ^791       

gaaaaccgcc tcgcggtgat ggtgctgcgt tggagtgacg gcagttatct ggaagatcag gatatgtggc ggatgagcgg cattttccgt gacgtctcgt 
^801       ^811       ^821       ^831       ^841       ^851       ^861       ^871       ^881       ^891       

tgctgcataa accgactaca caaatcagcg atttccatgt tgccactcgc tttaatgatg atttcagccg cgctgtactg gaggctgaag ttcagatgtg 
^901       ^911       ^921       ^931       ^941       ^951       ^961       ^971       ^981       ^991       

cggcgagttg cgtgactacc tacgggtaac agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt tcggcggtga aattatcgat 
^1001      ^1011      ^1021      ^1031      ^1041      ^1051      ^1061      ^1071      ^1081      ^1091      

gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg 
^1101      ^1111      ^1121      ^1131      ^1141      ^1151      ^1161      ^1171      ^1181      ^1191      

aactgcacac cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat tgaaaatggt ctgctgctgc tgaacggcaa 
^1201      ^1211      ^1221      ^1231      ^1241      ^1251      ^1261      ^1271      ^1281      ^1291      

gccgttgctg attcgaggcg ttaaccgtca cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc aggatatcct gctgatgaag 
^1301      ^1311      ^1321      ^1331      ^1341      ^1351      ^1361      ^1371      ^1381      ^1391      

cagaacaact ttaacgccgt gcgctgttcg cattatccga accatccgct gtggtacacg ctgtgcgacc gctacggcct gtatgtggtg gatgaagcca 
^1401      ^1411      ^1421      ^1431      ^1441      ^1451      ^1461      ^1471      ^1481      ^1491      

atattgaaac ccacggcatg gtgccaatga atcgtctgac cgatgatccg cgctggctac cggcgatgag cgaacgcgta acgcgaatgg tgcagcgcga 
^1501      ^1511      ^1521      ^1531      ^1541      ^1551      ^1561      ^1571      ^1581      ^1591      

tcgtaatcac ccgagtgtga tcatctggtc gctggggaat gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc tgtcgatcct 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

tcccgcccgg tgcagtatga aggcggcgga gccgacacca cggccaccga tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

ctgtgccgaa atggtccatc aaaaaatggc tttcgctacc tggagagacg cgcccgctga tcctttgcga atacgcccac gcgatgggta acagtcttgg 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

cggtttcgct aaatactggc aggcgtttcg tcagtatccc cgtttacagg gcggcttcgt ctgggactgg gtggatcagt cgctgattaa atatgatgaa 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

aacggcaacc cgtggtcggc ttacggcggt gattttggcg atacgccgaa cgatcgccag ttctgtatga acggtctggt ctttgccgac cgcacgccgc 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

atccagcgct gacggaagca aaacaccagc agcagttttt ccagttccgt ttatccgggc aaaccatcga agtgaccagc gaatacctgt tccgtcatag 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

cgataacgag ctcctgcact ggatggtggc gctggatggt aagccgctgg caagcggtga agtgcctctg gatgtcgctc cacaaggtaa acagttgatt 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

gaactgcctg aactaccgca gccggagagc gccgggcaac tctggctcac agtacgcgta gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

tcagcgcctg gcagcagtgg cgtctggcgg aaaacctcag tgtgacgctc cccgccgcgt cccacgccat cccgcatctg accaccagcg aaatggattt 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

ttgcatcgag ctgggtaata agcgttggca atttaaccgc cagtcaggct ttctttcaca gatgtggatt ggcgataaaa aacaactgct gacgccgctg 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

cgcgatcagt tcacccgtgc accgctggat aacgacattg gcgtaagtga agcgacccgc attgacccta acgcctgggt cgaacgctgg aaggcggcgg 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

gccattacca ggccgaagca gcgttgttgc agtgcacggc agatacactt gctgatgcgg tgctgattac gaccgctcac gcgtggcagc atcaggggaa 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

aaccttattt atcagccgga aaacctaccg gattgatggt agtggtcaaa tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

cggattggcc tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta gggccgcaag aaaactatcc cgaccgcctt actgccgcct 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

gttttgaccg ctgggatctg ccattgtcag acatgtatac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg acgcgcgaat tgaattatgg 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

cccacaccag tggcgcggcg acttccagtt caacatcagc cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc ccgtcagtat cggcggaatt acagctgagc gccggtcgct 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

accattacca gttggtctgg tgtcaaaaat aataataacc gggcaggcca tgtctgcccg tatttcgcgt aaggaaatcc attatgtact atttaaaaaa 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

cacaaacttt tggatgttcg gtttattctt tttcttttac ttttttatca tgggagccta cttcccgttt ttcccgattt ggctacatga catcaaccat 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

atcagcaaaa gtgatacggg tattattttt gccgctattt ctctgttctc gctattattc caaccgctgt ttggtctgct ttctgacaaa ctcggcctcg 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

actctaggcg gccgcctcga gtgatttctt tttgtggtaa aaatcttagt tcatgtgaag aaatttcatg tgaatgtttt agctatcaaa cagtactgtc 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

acctactcat acacaaaact gcctcccaaa gacttttccc aggtccctcg tatcaaaaca ttaagagtat actcgagatg gagaaaaaaa tcactggata 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt 
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371      ^4381      ^4391      

acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggtgcta cgcctgaata agtgataata 
^4401      ^4411      ^4421      ^4431      ^4441      ^4451      ^4461      ^4471      ^4481      ^4491      

agcggatgaa tggcagaaat tcgccggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta 
^4501      ^4511      ^4521      ^4531      ^4541      ^4551      ^4561      ^4571      ^4581      ^4591      

aggtaaatat aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg aatgggagca 
^4601      ^4611      ^4621      ^4631      ^4641      ^4651      ^4661      ^4671      ^4681      ^4691      

gtggtggaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca 
^4701      ^4711      ^4721      ^4731      ^4741      ^4751      ^4761      ^4771      ^4781      ^4791      

aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg 
^4801      ^4811      ^4821      ^4831      ^4841      ^4851      ^4861      ^4871      ^4881      ^4891      

ctatttacac cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg cataacagtt ataatcataa 
^4901      ^4911      ^4921      ^4931      ^4941      ^4951      ^4961      ^4971      ^4981      ^4991      

catactgttt tttcttactc cacacaggca tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt 
^5001      ^5011      ^5021      ^5031      ^5041      ^5051      ^5061      ^5071      ^5081      ^5091      

aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc 
^5101      ^5111      ^5121      ^5131      ^5141      ^5151      ^5161      ^5171      ^5181      ^5191      

cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 
^5201      ^5211      ^5221      ^5231      ^5241      ^5251      ^5261      ^5271      ^5281      ^5291      

caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgcctagag ggccctattc tatagtgtca 
^5301      ^5311      ^5321      ^5331      ^5341      ^5351      ^5361      ^5371      ^5381      ^5391      

cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 
^5401      ^5411      ^5421      ^5431      ^5441      ^5451      ^5461      ^5471      ^5481      ^5491      

gccactccca ctgtcctttc ctaataa
^5501      ^5511      ^5521

Credibility of mRNA sequence:

reverse_engineered_sequence_and_should_match_experiment_except_3UTR

The name of the plasmid:
pbetaGAL/HIAP2(1073-1222)/CAT The name of the promoter used to express this mRNA:
CMV

Description of the plasmid (facultative for promoter-less plasmid records):
Plasmid containing part of human c-IAP1 5' UTR (nt from -150 to nt -1 of the original sequence) inserted between beta galactosidase and chloramphenicol acetyltransferase reporter genes.

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

no

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

not tested

A promoter reported in cDNA corresponding to IRES sequence:

no (not convincing)

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:
not_tested

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

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:

Homo sapiens HEK 293T (ATCC 293tsA1609neo)

The abbreviated name of the donor gene or virus from which this IRES was excised and inserted into the plasmid:
c-IAP1

The origin of IRES in the plasmid:

nuclear

The donor organism of the IRES segment:

Homo sapiens Jurkat

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

cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc 
^1         ^11        ^21        ^31        ^41        ^51        ^61        ^71        ^81        ^91        

ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 
^101       ^111       ^121       ^131       ^141       ^151       ^161       ^171       ^181       ^191       

cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg 
^201       ^211       ^221       ^231       ^241       ^251       ^261       ^271       ^281       ^291       

gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 
^301       ^311       ^321       ^331       ^341       ^351       ^361       ^371       ^381       ^391       

aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 
^401       ^411       ^421       ^431       ^441       ^451       ^461       ^471       ^481       ^491       

ccattgacgt caatgggtgg actatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac 
^501       ^511       ^521       ^531       ^541       ^551       ^561       ^571       ^581       ^591       

ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 
^601       ^611       ^621       ^631       ^641       ^651       ^661       ^671       ^681       ^691       

tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 
^701       ^711       ^721       ^731       ^741       ^751       ^761       ^771       ^781       ^791       

ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct 
^801       ^811       ^821       ^831       ^841       ^851       ^861       ^871       ^881       ^891       

ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta ccgagctcgg atccactagt 
^901       ^911       ^921       ^931       ^941       ^951       ^961       ^971       ^981       ^991       

aacggccgcc agtgtgctgg aattctgcag atatccatca cactggcggc cgcaattccc ggggatcgaa agagcctgct aaagcaaaaa agaagtcacc 
^1001      ^1011      ^1021      ^1031      ^1041      ^1051      ^1061      ^1071      ^1081      ^1091      

atgtcgttta ctttgaccaa caagaacgtg attttcgttg ccggtctggg aggcattggt ctggacacca gcaaggagct gctcaagcgc gatcccgtcg 
^1101      ^1111      ^1121      ^1131      ^1141      ^1151      ^1161      ^1171      ^1181      ^1191      

ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg 
^1201      ^1211      ^1221      ^1231      ^1241      ^1251      ^1261      ^1271      ^1281      ^1291      

caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc 
^1301      ^1311      ^1321      ^1331      ^1341      ^1351      ^1361      ^1371      ^1381      ^1391      

gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca 
^1401      ^1411      ^1421      ^1431      ^1441      ^1451      ^1461      ^1471      ^1481      ^1491      

atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt 
^1501      ^1511      ^1521      ^1531      ^1541      ^1551      ^1561      ^1571      ^1581      ^1591      

tgatggcgtt aactcggcgt ttcatctgtg gtgcaacggg cgctgggtcg gttacggcca ggacagtcgt ttgccgtctg aatttgacct gagcgcattt 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

ttacgcgccg gagaaaaccg cctcgcggtg atggtgctgc gttggagtga cggcagttat ctggaagatc aggatatgtg gcggatgagc ggcattttcc 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

gtgacgtctc gttgctgcat aaaccgacta cacaaatcag cgatttccat gttgccactc gctttaatga tgatttcagc cgcgctgtac tggaggctga 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

agttcagatg tgcggcgagt tgcgtgacta cctacgggta acagtttctt tatggcaggg tgaaacgcag gtcgccagcg gcaccgcgcc tttcggcggt 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

gaaattatcg atgagcgtgg tggttatgcc gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac tgtggagcgc cgaaatcccg aatctctatc 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

gtgcggtggt tgaactgcac accgccgacg gcacgctgat tgaagcagaa gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg gtctgctgct 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

gctgaacggc aagccgttgc tgattcgagg cgttaaccgt cacgagcatc atcctctgca tggtcaggtc atggatgagc agacgatggt gcaggatatc 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

ctgctgatga agcagaacaa ctttaacgcc gtgcgctgtt cgcattatcc gaaccatccg ctgtggtaca cgctgtgcga ccgctacggc ctgtatgtgg 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

tggatgaagc caatattgaa acccacggca tggtgccaat gaatcgtctg accgatgatc cgcgctggct accggcgatg agcgaacgcg taacgcgaat 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

ggtgcagcgc gatcgtaatc acccgagtgt gatcatctgg tcgctgggga atgaatcagg ccacggcgct aatcacgacg cgctgtatcg ctggatcaaa 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

tctgtcgatc cttcccgccc ggtgcagtat gaaggcggcg gagccgacac cacggccacc gatattattt gcccgatgta cgcgcgcgtg gatgaagacc 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

agcccttccc ggctgtgccg aaatggtcca tcaaaaaatg gctttcgcta cctggagaga cgcgcccgct gatcctttgc gaatacgccc acgcgatggg 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

taacagtctt ggcggtttcg ctaaatactg gcaggcgttt cgtcagtatc cccgtttaca gggcggcttc gtctgggact gggtggatca gtcgctgatt 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

aaatatgatg aaaacggcaa cccgtggtcg gcttacggcg gtgattttgg cgatacgccg aacgatcgcc agttctgtat gaacggtctg gtctttgccg 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

accgcacgcc gcatccagcg ctgacggaag caaaacacca gcagcagttt ttccagttcc gtttatccgg gcaaaccatc gaagtgacca gcgaatacct 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

gttccgtcat agcgataacg agctcctgca ctggatggtg gcgctggatg gtaagccgct ggcaagcggt gaagtgcctc tggatgtcgc tccacaaggt 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

aaacagttga ttgaactgcc tgaactaccg cagccggaga gcgccgggca actctggctc acagtacgcg tagtgcaacc gaacgcgacc gcatggtcag 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

aagccgggca catcagcgcc tggcagcagt ggcgtctggc ggaaaacctc agtgtgacgc tccccgccgc gtcccacgcc atcccgcatc tgaccaccag 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

cgaaatggat ttttgcatcg agctgggtaa taagcgttgg caatttaacc gccagtcagg ctttctttca cagatgtgga ttggcgataa aaaacaactg 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

ctgacgccgc tgcgcgatca gttcacccgt gcaccgctgg ataacgacat tggcgtaagt gaagcgaccc gcattgaccc taacgcctgg gtcgaacgct 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

ggaaggcggc gggccattac caggccgaag cagcgttgtt gcagtgcacg gcagatacac ttgctgatgc ggtgctgatt acgaccgctc acgcgtggca 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

gcatcagggg aaaaccttat ttatcagccg gaaaacctac cggattgatg gtagtggtca aatggcgatt accgttgatg ttgaagtggc gagcgataca 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

ccgcatccgg cgcggattgg cctgaactgc cagctggcgc aggtagcaga gcgggtaaac tggctcggat tagggccgca agaaaactat cccgaccgcc 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

ttactgccgc ctgttttgac cgctgggatc tgccattgtc agacatgtat accccgtacg tcttcccgag cgaaaacggt ctgcgctgcg ggacgcgcga 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

attgaattat ggcccacacc agtggcgcgg cgacttccag ttcaacatca gccgctacag tcaacagcaa ctgatggaaa ccagccatcg ccatctgctg 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

cacgcggaag aaggcacatg gctgaatatc gacggtttcc atatggggat tggtggcgac gactcctgga gcccgtcagt atcggcggaa ttacagctga 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

gcgccggtcg ctaccattac cagttggtct ggtgtcaaaa ataataataa ccgggcaggc catgtctgcc cgtatttcgc gtaaggaaat ccattatgta 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

ctatttaaaa aacacaaact tttggatgtt cggtttattc tttttctttt acttttttat catgggagcc tacttcccgt ttttcccgat ttggctacat 
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371      ^4381      ^4391      

gacatcaacc atatcagcaa aagtgatacg ggtattattt ttgccgctat ttctctgttc tcgctattat tccaaccgct gtttggtctg ctttctgaca 
^4401      ^4411      ^4421      ^4431      ^4441      ^4451      ^4461      ^4471      ^4481      ^4491      

aactcggcct cgactctagg cggccgcctc gagtgatttc tttttgtggt aaaaatctta gttcatgtga agaaatttca tgtgaatgtt ttagctatca 
^4501      ^4511      ^4521      ^4531      ^4541      ^4551      ^4561      ^4571      ^4581      ^4591      

aacagtactg tcacctactc atacacaaaa ctgcctccca aagacttttc ccaggtccct cgtatcaaaa cattaagagt atactcgaga tggagaaaaa 
^4601      ^4611      ^4621      ^4631      ^4641      ^4651      ^4661      ^4671      ^4681      ^4691      

aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt 
^4701      ^4711      ^4721      ^4731      ^4741      ^4751      ^4761      ^4771      ^4781      ^4791      

cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc 
^4801      ^4811      ^4821      ^4831      ^4841      ^4851      ^4861      ^4871      ^4881      ^4891      

cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc 
^4901      ^4911      ^4921      ^4931      ^4941      ^4951      ^4961      ^4971      ^4981      ^4991      

gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg 
^5001      ^5011      ^5021      ^5031      ^5041      ^5051      ^5061      ^5071      ^5081      ^5091      

tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt 
^5101      ^5111      ^5121      ^5131      ^5141      ^5151      ^5161      ^5171      ^5181      ^5191      

tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat 
^5201      ^5211      ^5221      ^5231      ^5241      ^5251      ^5261      ^5271      ^5281      ^5291      

gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggtgc tacgcctgaa 
^5301      ^5311      ^5321      ^5331      ^5341      ^5351      ^5361      ^5371      ^5381      ^5391      

taagtgataa taagcggatg aatggcagaa attcgccgga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga 
^5401      ^5411      ^5421      ^5431      ^5441      ^5451      ^5461      ^5471      ^5481      ^5491      

tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct atggaactga 
^5501      ^5511      ^5521      ^5531      ^5541      ^5551      ^5561      ^5571      ^5581      ^5591      

tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat 
^5601      ^5611      ^5621      ^5631      ^5641      ^5651      ^5661      ^5671      ^5681      ^5691      

tctactcctc caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt aatagaactc 
^5701      ^5711      ^5721      ^5731      ^5741      ^5751      ^5761      ^5771      ^5781      ^5791      

ttgcttgctt tgctatttac accacaaagg aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta ggcataacag 
^5801      ^5811      ^5821      ^5831      ^5841      ^5851      ^5861      ^5871      ^5881      ^5891      

ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt 
^5901      ^5911      ^5921      ^5931      ^5941      ^5951      ^5961      ^5971      ^5981      ^5991      

tgtaaagggg ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 
^6001      ^6011      ^6021      ^6031      ^6041      ^6051      ^6061      ^6071      ^6081      ^6091      

ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 
^6101      ^6111      ^6121      ^6131      ^6141      ^6151      ^6161      ^6171      ^6181      ^6191      

tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgcctag agggccctat 
^6201      ^6211      ^6221      ^6231      ^6241      ^6251      ^6261      ^6271      ^6281      ^6291      

tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 
^6301      ^6311      ^6321      ^6331      ^6341      ^6351      ^6361      ^6371      ^6381      ^6391      

accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 
^6401      ^6411      ^6421      ^6431      ^6441      ^6451      ^6461      ^6471      ^6481      ^6491      

ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 
^6501      ^6511      ^6521      ^6531      ^6541      ^6551      ^6561      ^6571      ^6581      ^6591      

tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 
^6601      ^6611      ^6621      ^6631      ^6641      ^6651      ^6661      ^6671      ^6681      ^6691      

gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt 
^6701      ^6711      ^6721      ^6731      ^6741      ^6751      ^6761      ^6771      ^6781      ^6791      

tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 
^6801      ^6811      ^6821      ^6831      ^6841      ^6851      ^6861      ^6871      ^6881      ^6891      

gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat 
^6901      ^6911      ^6921      ^6931      ^6941      ^6951      ^6961      ^6971      ^6981      ^6991      

tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag 
^7001      ^7011      ^7021      ^7031      ^7041      ^7051      ^7061      ^7071      ^7081      ^7091      

gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 
^7101      ^7111      ^7121      ^7131      ^7141      ^7151      ^7161      ^7171      ^7181      ^7191      

ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 
^7201      ^7211      ^7221      ^7231      ^7241      ^7251      ^7261      ^7271      ^7281      ^7291      

atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 
^7301      ^7311      ^7321      ^7331      ^7341      ^7351      ^7361      ^7371      ^7381      ^7391      

cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 
^7401      ^7411      ^7421      ^7431      ^7441      ^7451      ^7461      ^7471      ^7481      ^7491      

ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 
^7501      ^7511      ^7521      ^7531      ^7541      ^7551      ^7561      ^7571      ^7581      ^7591      

aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 
^7601      ^7611      ^7621      ^7631      ^7641      ^7651      ^7661      ^7671      ^7681      ^7691      

ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 
^7701      ^7711      ^7721      ^7731      ^7741      ^7751      ^7761      ^7771      ^7781      ^7791      

ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 
^7801      ^7811      ^7821      ^7831      ^7841      ^7851      ^7861      ^7871      ^7881      ^7891      

gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 
^7901      ^7911      ^7921      ^7931      ^7941      ^7951      ^7961      ^7971      ^7981      ^7991      

atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 
^8001      ^8011      ^8021      ^8031      ^8041      ^8051      ^8061      ^8071      ^8081      ^8091      

ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 
^8101      ^8111      ^8121      ^8131      ^8141      ^8151      ^8161      ^8171      ^8181      ^8191      

gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 
^8201      ^8211      ^8221      ^8231      ^8241      ^8251      ^8261      ^8271      ^8281      ^8291      

acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 
^8301      ^8311      ^8321      ^8331      ^8341      ^8351      ^8361      ^8371      ^8381      ^8391      

ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 
^8401      ^8411      ^8421      ^8431      ^8441      ^8451      ^8461      ^8471      ^8481      ^8491      

tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 
^8501      ^8511      ^8521      ^8531      ^8541      ^8551      ^8561      ^8571      ^8581      ^8591      

gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 
^8601      ^8611      ^8621      ^8631      ^8641      ^8651      ^8661      ^8671      ^8681      ^8691      

acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 
^8701      ^8711      ^8721      ^8731      ^8741      ^8751      ^8761      ^8771      ^8781      ^8791      

gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 
^8801      ^8811      ^8821      ^8831      ^8841      ^8851      ^8861      ^8871      ^8881      ^8891      

atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 
^8901      ^8911      ^8921      ^8931      ^8941      ^8951      ^8961      ^8971      ^8981      ^8991      

aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 
^9001      ^9011      ^9021      ^9031      ^9041      ^9051      ^9061      ^9071      ^9081      ^9091      

gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 
^9101      ^9111      ^9121      ^9131      ^9141      ^9151      ^9161      ^9171      ^9181      ^9191      

taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 
^9201      ^9211      ^9221      ^9231      ^9241      ^9251      ^9261      ^9271      ^9281      ^9291      

cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 
^9301      ^9311      ^9321      ^9331      ^9341      ^9351      ^9361      ^9371      ^9381      ^9391      

tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 
^9401      ^9411      ^9421      ^9431      ^9441      ^9451      ^9461      ^9471      ^9481      ^9491      

atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 
^9501      ^9511      ^9521      ^9531      ^9541      ^9551      ^9561      ^9571      ^9581      ^9591      

tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 
^9601      ^9611      ^9621      ^9631      ^9641      ^9651      ^9661      ^9671      ^9681      ^9691      

aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 
^9701      ^9711      ^9721      ^9731      ^9741      ^9751      ^9761      ^9771      ^9781      ^9791      

tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 
^9801      ^9811      ^9821      ^9831      ^9841      ^9851      ^9861      ^9871      ^9881      ^9891      

cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 
^9901      ^9911      ^9921      ^9931      ^9941      ^9951      ^9961      ^9971      ^9981      ^9991      

attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 
^10001     ^10011     ^10021     ^10031     ^10041     ^10051     ^10061     ^10071     ^10081     ^10091     

tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 
^10101     ^10111     ^10121     ^10131     ^10141     ^10151     ^10161     ^10171     ^10181     ^10191     

gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 
^10201     ^10211     ^10221     ^10231     ^10241     ^10251     ^10261     ^10271     ^10281     ^10291     

gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 
^10301     ^10311     ^10321     ^10331     ^10341     ^10351     ^10361     ^10371     ^10381     ^10391     

tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 
^10401     ^10411     ^10421     ^10431     ^10441     ^10451     ^10461     ^10471     ^10481     ^10491     

aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaagg
^10501     ^10511     ^10521     ^10531     ^10541     ^10551     ^10561     ^10571

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

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: Václav Vopálenský	Reviewed by: Martin Mokrejš

The abbreviated name of this ORF/gene:
LacZ

The description of the protein encoded in this ORF:
beta galactosidase

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:

not tested

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

189-3332

ORF

ORF position:

2

Version: `1`	Last change: `2008-10-27 23:35:24`
Originaly submitted by: Václav Vopálenský	Reviewed by: Martin Mokrejš

The abbreviated name of this ORF/gene:
CAT

The description of the protein encoded in this ORF:
chloramphenicol acetyltransferase

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:

not tested

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

3778-4437

Remarks:

Unclear results of ribonuclease protection assay because of not good quality of Figure 2B. There are some
minor bands clearly visible on the PAA gel (Figure 2B).

Citations:

Warnakulasuriyarachchi D., Cerquozzi S., Cheung H. H., Holcik M. (2004) Translational induction of the inhibitor of apoptosis protein HIAP2 during endoplasmic reticulum stress attenuates cell death and is mediated via an inducible internal ribosome entry site element. J. Biol. Chem. 279(17):17148-17157

IRESs:

IRES:

Version: `1`	Last change: `2008-10-23 13:15:19`
Originaly submitted by: Václav Vopálenský	Reviewed by: Martin Mokrejš
The IRES name: `c-IAP1_-150/-1`	The functional status of IRES: `functional`

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

3622-3771

How IRES boundaries were determined:
experimentally_determined

5'-end of IRES relative to last base of the STOP codon of the upstream ORF: `290`	3'-end of IRES relative to last base of the STOP codon of the upstream ORF: `439`
5'-end of IRES relative to first base of the START codon of the downstream ORF: `-156`	3'-end of IRES relative to first base of the START codon of the downstream ORF: `-7`

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

tgatttcttt ttgtggtaaa aatcttagtt catgtgaaga aatttcatgt gaatgtttta gctatcaaac agtactgtca cctactcata cacaaaactg 
^3622      ^3632      ^3642      ^3652      ^3662      ^3672      ^3682      ^3692      ^3702      ^3712      

cctcccaaag acttttccca ggtccctcgt atcaaaacat taagagtata
^3722      ^3732      ^3742      ^3752      ^3762

Remarks:

c-IAP1_-150/-1 IRES represents part of the human c-IAP1 5' UTR (nt from -150 to nt -1).

Citations:

The translation experiments:

Translation results:

IRESite Translation Id: 578

Version: `7`	Last change: `2008-10-27 23:35:24`
Originaly submitted by: Václav Vopálenský	Reviewed by: Martin Mokrejš

The translation method used to study IRES function:
in vivo

The organism used for translation:

Homo sapiens HEK 293T (ATCC 293tsA1609neo)

The temperature (in degrees of Celsia):
37

The relative translation efficiency in % of this IRES:

156.100

Name of IRES used as the positive control:

cIAP_-1222/-1

Name of the plasmid used as the positive control. `pbetaGAL/HIAP2(-1222/-1)/CAT`
`IRESite Id` of the plasmid used as positive control. `524`
The relative translation efficiency in % of the positive control: `100.000`
The size (length) of intercistronic region in the positive control: `1565`

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 IRES activity was determined in p86-cotrasfected HEK293T cells because the caspase-mediated
cleavage fragment of p97/DAP5/NAT1, p86, selectively enhances translation of c-IAP1 IRES element both in vivo
and in vitro in a manner that is independent of the translation of the upstream cistron
(Warnakulasuriyarachchi et al., 2004).
Translational data are derived from Fig. 5A.

Similar results were obtained in vitro using rabbit reticulocyte lysate, where is possible to prove the
production of CAT protein on SDS-PAGE gel, compared with the bicistronic construct with shorter the 5' UTR
insert (pbetaGAL/HIAP2(1142-1222)/CAT). See Figure 5B.

Citations:

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