LOCUS Exported 6071 bp ds-DNA circular SYN 16-FEB-2017 DEFINITION E1 Shuttle plasmid for constructing adenovirus vectors with the AdenoQuick2.0 system. ACCESSION . VERSION . KEYWORDS pAd1127 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6071) AUTHORS . TITLE . JOURNAL . REFERENCE 2 (bases 1 to 6071) AUTHORS OD260 Inc TITLE Direct Submission JOURNAL Exported Thursday, Feb 16, 2017 from SnapGene 3.3.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6071 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 5..107 /label=Ad5 left ITR /note="Ad5 left ITR" /note="ITR" regulatory 56..61 /regulatory_class="GC_signal" /note="SP1" /note="SP1 binding site - promotes bidirectional transcription from ITR in vitro (Hatfield and Hearing, Virology 1991; 184:265-76)" promoter 68..76 /note="ATF binding site" regulatory 68..73 /regulatory_class="promoter" /note="ATF" /note="ATF binding site -" regulatory 100..105 /regulatory_class="promoter" /note="ATF" /note="ATF binding site -" enhancer 159..165 /note="Enhancer I" /note="Enhancer I element" regulatory 174..179 /regulatory_class="promoter" /note="ATF" /note="ATF binding site -" misc_signal 194..454 /label=Ad5 Packaging Signal/Enhancers /note="Ad5 PSI" /note="Contains all seven A-repeats" enhancer 203..209 /note="Enhancer I" /note="enhancer I element" promoter 216..222 /note="E2F" /note="E2F binding site" enhancer 233..239 /note="Enhancer I" /note="Enhancer I element" misc_feature 244..252 /function="Ad5 genome packaging" /note="A1" /note="- A1 repeat - consensus sequence: 5'-TTTG-N8-CG-3' - Ostapchuk and Hearing (2005) J Cell Biochem. 96:25-35" misc_feature 264..273 /function="Ad5 genome packaging" /note="A2" /note="- A2 repeat - consensus sequence: 5'-TTTG-N8-CG-3' - Ostapchuk and Hearing (2005) J Cell Biochem. 96:25-35" promoter 279..285 /note="E2F" /note="E2F binding site" enhancer 301..307 /note="Enhancer I" /note="enhancer I element" misc_feature 306..315 /function="Ad5 DNA packaging" /note="A3" /note="- A3 repeat - Ostapchuk and Hearing (2005) J Cell Biochem. 96:25-35" misc_feature 317..325 /function="Ad5 DNA packaging" /note="A4" /note="- A4 repeat - Ostapchuk and Hearing (2005) J Cell Biochem. 96:25-35" misc_feature 341..350 /function="Ad5 DNA packaging" /note="A5" /note="- A5 repeat - consensus sequence: 5'-TTTG-N8-CG-3' - Ostapchuk and Hearing (2005) J Cell Biochem. 96:25-35" misc_feature 363..371 /function="Ad5 DNA packaging" /note="A6" /note="- A6 repeat - consensus sequence: 5'-TTTG-N8-CG-3' - Ostapchuk and Hearing (2005) J Cell Biochem. 96:25-35" misc_feature 371..379 /function="Ad5 DNA packaging" /note="A7" /note="- A7 repeat - Ostapchuk and Hearing (2005) J Cell Biochem. 96:25-35" regulatory 460..465 /regulatory_class="promoter" /note="ATF" /note="ATF binding site -" gene 472..1636 /product="- The primary E1A transcript is processed by differential splicing to yield 5 distinct mRNAs: 13S, 12S, 11S, 10S, and 9 S, which respectively code for proteins with 289 residues (289R), 243R, 217R, 171R, and 55R, all of which are detectable in vivo with the exception of the 9S product which has only been detected in vitro. (http://publish.uwo.ca/~jmymryk/E1A/intro.html) - The 13S and 12S E1A mRNAs are the most abundant at early times during infection. The 9S nRNA is the most abundant at late times. The 11S and 10S are minor species that become more abundant at late times after the infection. (http://publish.uwo.ca/~jmymryk/E1A/intro.html)" /function="- The E1A proteins, especially the 289R, augment E1A gene transcription to high levels, and also stimulate promoter activities of other viral genes, including the promoter for the E1B, E1A, E3, and E4 early genes, as well as the major late promoter (Yoshida et al, 1995, The molecular Repertoire of Adenovirus III, pp113-130) - E1A proteins reprogram cell growth to provide an optimal environment for viral replication. They interact with a variety of cellular proteins, including transcriptional co-activators such as the CREB-binding protein (CBP) and p300, the p300/CBP Associated Factor (pCAF), and the transcriptional co-repressors CtBP and BS69, the TAT-binding (TBP) protein component of the general transcriptional machinery, pRB and related p130 and p107, the cyclin-dependent kinase inhibitor p27, SUMO conjugase UBC9 and proteasome components (Avvakumov et al, 2004, Virology 329: 477-492)" /note="Ad5 E1A" /note="- first gene expressed after infection of the host cell - Ad5 E1A = Ad2 E1A" regulatory 472..479 /regulatory_class="TATA_box" /gene="E1A" /note="E1A TATA Box" CDS join(564..1116,1233..1549) /codon_start=1 /gene="E1A" /product="E1A 289R = 32 kDa protein" /label=Ad5 E1a /note="Ad5 E1A 289R" /note="- derived from the E1A 13S mRNA - The E1A 289R protein has 4 conserved regions (CR1, CR2, CR3, and CR4), based on the alignment of E1A aa sequences from 34 human and simian adenovirus strains (Avvakumov et al, Virology 2004, 329: 477-492). However there are conserved motifs outside of these conserved regions, that are important for the binding of E1A to other proteins. For instance, Ad5 E1A aa residues 19-28 LLDQLIEEVL are important for binding to p300/CBP (Wang et al, 1993, J Virol 67:476-488)" /db_xref="GI:33465831" /protein_id="AAQ19284.1" /translation="MRHIICHGGVITEEMAASLLDQLIEEVLADNLPPPSHFEPPTLHE LYDLDVTAPEDPNEEAVSQIFPDSVMLAVQEGIDLLTFPPAPGSPEPPHLSRQPEQPEQ RALGPVSMPNLVPEVIDLTCHEAGFPPSDDEDEEGEEFVLDYVEHPGHGCRSCHYHRRN TGDPDIMCSLCYMRTCGMFVYSPVSEPEPEPEPEPEPARPTRRPKMAPAILRRPTSPVS RECNSSTDSCDSGPSNTPPEIHPVVPLCPIKPVAVRVGGRRQAVECIEDLLNEPGQPLD LSCKRPRP" CDS join(564..978,1233..1548) /codon_start=1 /gene="E1A" /product="E1A 243R = 27 kDa protein" /note="Ad5 E1A 243R" /note="derived from the E1A 12S mRNA" /db_xref="GI:33465832" /protein_id="AAQ19285.1" /translation="MRHIICHGGVITEEMAASLLDQLIEEVLADNLPPPSHFEPPTLHE LYDLDVTAPEDPNEEAVSQIFPDSVMLAVQEGIDLLTFPPAPGSPEPPHLSRQPEQPEQ RALGPVSMPNLVPEVIDLTCHEAGFPPSDDEDEEGPVSEPEPEPEPEPEPARPTRRPKM APAILRRPTSPVSRECNSSTDSCDSGPSNTPPEIHPVVPLCPIKPVAVRVGGRRQAVEC IEDLLNEPGQPLDLSCKRPRP" protein_bind 618..647 /bound_moiety="CREB-binding protein (CBP) and p300" /note="CBP" /note="Avvakumov et al, 2004, Virology 329:477-492 " misc_feature 687..779 /note="CR1" /note="- E1A Conserved Region 1 in human and simian adenovirus serotypes - overlaps with the weakest of two Rb-binding sites (HFEPPTLHELYDLDVTAP, aa 37-54), the strongest site being located in CR2 - contains one of three binding sites for CREB-binding protein (CBP)/p300: FPDSVML (consensus FXE/DXXXL). The two other CBP binding sites are located at the N-terminal end of E1A. (Avvakumov et al, Virology 2004, 329:477-492)" protein_bind 759..779 /bound_moiety="TRAM motif located in CH3 region of the CREB-binding protein (CBP)" /note="CBP" /note="O'Connor et al, 1999. J. Virol 73: 3574-3581" misc_feature 906..974 /gene="E1A" /function="Mediates binding of E1A proteins to pRB family proteins (pRB, p107, p130), dissociating them from the E2F transcription factor. E2F can then activate cellular genes involved in cell cycle and DNA synthesis and also the E2 viral genes." /note="CR2" /note="- E1A Conserved Region 2 among human and simian adenovirus serotypes - a.p. 122-126: consensus sequence LXCXE (where X is variable), which is the strongest of 2 binding sites for Rb protein - a.p. 113-117: consensus sequence PXLXP, which binds to transcriptional co-repressor BS69 (Avvakumov et al, Virology 2004, 329:477-92)." misc_feature 927..950 /note="D24" /note="- 24-bp deletion (= aa LTCHEAGF), which encompasses the Rb-binding consensus site LXCXE (where X is variable). LXCXE is absolutely conserved in all 34 E1A sequences analyzed by Avvakumov (Virology 2004, 329: 477-492). In Ad5, the phenylalanine residue located 3 aa downstream from the LXCXE motif could interact with a hydrophobic pocket on Rb. - the 24-bp deletion helps targeting oncolytic adenoviruses to tumor cells characterized by abnormal Rb control (Fueyo et al, Oncogene 200, 19:2-12)." regulatory 951..956 /regulatory_class="GC_signal" /note="SP1" /note="SP1 binding site - promotes bidirectional transcription from ITR in vitro (Hatfield and Hearing, Virology 1991; 184:265-76)" intron 979..1232 /gene="E1A" /note="E1A 12S" misc_feature join(993..1116,1233..1252) /note="CR3" /note="- E1A Conserved Region 3 in human and simian adenovirus serotypes - Cysteins at positions 154, 157, 171, and 174 form the zinc finger and are absolutely conserved in 34 human and simian adenovirus strains analyzed by Avvakumov et al (Virology 2004, 329:477-92)." intron 1117..1232 /gene="E1A" /note="E1A 13S" misc_feature 1397..1543 /note="CR4" /note="- E1A Conserved Region 4 in human and simian adenovirus serotypes - Contains 2 binding sites for the transcriptional co-repressor CtBP - Overlaps with the C-terminal Nuclear Localization Signal KRPRP - The role of most of the conserved aa in CR4 is unknown (Avvakumov et al, Virology 2004, 329:477-92)" misc_signal 1532..1546 /note="E1A NLS" /note="- conventional, constitutively active E1A nuclear localization signal - There is a second, non-conventional, developmentally-regulated NLS with the consensus: FV(X)7-20MXSLXYM(X)4MF, located in CR3, thus unique for the larger 289R protein" regulatory 1615..1620 /regulatory_class="polyA_signal_sequence" /gene="E1A" /note="E1A polyA signal" regulatory 1658..1666 /regulatory_class="GC_signal" /function="Binding site for SP1 transcription factor" /note="SP1 binding site" /note="-The presence of the GC box and TATA box is sufficient for high level of E1B gene transcription (Parks et al, J. Virol. 1988, 62:54-67)." gene 1676..3513 /gene="E1B" /note="E1B gene" regulatory 1676..1682 /regulatory_class="TATA_box" /gene="E1B" /note="E1B TATA Box" CDS 1718..2248 /codon_start=1 /gene="E1B" /product="Ad5 E1B 19K (= 21K)" /function="homolog of Bcl2 that blocks apoptosis in infected cells" /label=Ad5 E1b 19K /note="Ad5 E1B 19K" /db_xref="GI:33465833" /protein_id="AAQ19286.1" /translation="MEAWECLEDFSAVRNLLEQSSNSTSWFWRFLWGSSQAKLVCRIKE DYKWEFEELLKSCGELFDSLNLGHQALFQEKVIKTLDFSTPGRAAAAVAFLSFIKDKWS EETHLSGGYLLDFLAMHLWRAVVRHKNRLLLLSSVRPAIIPTEEQQQQQEEARRRRQEQ SPWNPRAGLDPRE" CDS 2023..3513 /codon_start=1 /gene="E1B" /product="Ad5 E1B 55K" /function="- binds and inhibits p53" /label=Ad5 E1b 55K /note="Ad5 E1B 55K" /db_xref="GI:33465834" /protein_id="AAQ19287.1" /translation="MERRNPSERGVPAGFSGHASVESGCETQESPATVVFRPPGDNTDG GAAAAAGGSQAAAAGAEPMEPESRPGPSGMNVVQVAELYPELRRILTITEDGQGLKGVK RERGACEATEEARNLAFSLMTRHRPECITFQQIKDNCANELDLLAQKYSIEQLTTYWLQ PGDDFEEAIRVYAKVALRPDCKYKISKLVNIRNCCYISGNGAEVEIDTEDRVAFRCSMI NMWPGVLGMDGVVIMNVRFTGPNFSGTVFLANTNLILHGVSFYGFNNTCVEAWTDVRVR GCAFYCCWKGVVCRPKSRASIKKCLFERCTLGILSEGNSRVRHNVASDCGCFMLVKSVA VIKHNMVCGNCEDRASQMLTCSDGNCHLLKTIHVASHSRKAWPVFEHNILTRCSLHLGN RRGVFLPYQCNLSHTKILLEPESMSKVNLNGVFDMTMKIWKVLRYDETRTRCRPCECGG KHIRNQPVMLDVTEELRPDHLVLACTRAEFGSSDEDTD" regulatory 2663..2668 /regulatory_class="GC_signal" /note="SP1" /note="SP1 binding site - promotes bidirectional transcription from ITR in vitro (Hatfield and Hearing, Virology 1991; 184:265-76)" regulatory 2720..2725 /regulatory_class="GC_signal" /note="SP1" /note="SP1 binding site - promotes bidirectional transcription from ITR in vitro (Hatfield and Hearing, Virology 1991; 184:265-76)" regulatory 2894..2899 /regulatory_class="GC_signal" /note="SP1" /note="SP1 binding site - promotes bidirectional transcription from ITR in vitro (Hatfield and Hearing, Virology 1991; 184:265-76)" regulatory 3542..3547 /regulatory_class="GC_signal" /note="SP1" /note="SP1 binding site - promotes bidirectional transcription from ITR in vitro (Hatfield and Hearing, Virology 1991; 184:265-76)" regulatory 3555..3560 /regulatory_class="TATA_box" /note="pIX TATA box" /note="pIX TATA box" CDS 3613..4035 /codon_start=1 /gene="pIX" /product="Ad5 pIX" /label=Ad5 pIX /note="Ad5 pIX" /db_xref="GI:33465835" /protein_id="AAQ19288.1" /translation="MSTNSFDGSIVSSYLTTRMPPWAGVRQNVMGSSIDGRPVLPANST TLTYETVSGTPLETAASAAASAAAATARGIVTDFAFLSPLASSAASRSSARDDKLTALL AQLDSLTRELNVVSQQLLDLRQQVSALKASSPPNAV" rep_origin complement(4111..4699) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4874..5689) /codon_start=1 /product="Kanamycin-R" /label=Kanamycin-R /note="Kanamycin-R" /note="origin: Tn903" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" misc_signal 5752..5999 /label=Cos site /note="Cos site" /note="phage lambda COS site" ORIGIN 1 ttaacatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg 61 gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag tgtggcggaa 121 gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt 181 tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt 241 gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa actgaataag 301 aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggccg 361 cggggacttt gaccgtttac gtggagactc gcccaggtgt ttttctcagg tgttttccgc 421 gttccgggtc aaagttggcg ttttattatt atagtcagct gacgtgtagt gtatttatac 481 ccggtgagtt cctcaagagg ccactcttga gtgccagcga gtagagtttt ctcctccgag 541 ccgctccgac accgggactg aaaatgagac atattatctg ccacggaggt gttattaccg 601 aagaaatggc cgccagtctt ttggaccagc tgatcgaaga ggtactggct gataatcttc 661 cacctcctag ccattttgaa ccacctaccc ttcacgaact gtatgattta gacgtgacgg 721 cccccgaaga tcccaacgag gaggcggttt cgcagatttt tcccgactct gtaatgttgg 781 cggtgcagga agggattgac ttactcactt ttccgccggc gcccggttct ccggagccgc 841 ctcacctttc ccggcagccc gagcagccgg agcagagagc cttgggtccg gtttctatgc 901 caaaccttgt accggaggtg atcgatctta cctgccacga ggctggcttt ccacccagtg 961 acgacgagga tgaagagggt gaggagtttg tgttagatta tgtggagcac cccgggcacg 1021 gttgcaggtc ttgtcattat caccggagga atacggggga cccagatatt atgtgttcgc 1081 tttgctatat gaggacctgt ggcatgtttg tctacagtaa gtgaaaatta tgggcagtgg 1141 gtgatagagt ggtgggtttg gtgtggtaat ttttttttta atttttacag ttttgtggtt 1201 taaagaattt tgtattgtga tttttttaaa aggtcctgtg tctgaacctg agcctgagcc 1261 cgagccagaa ccggagcctg caagacctac ccgccgtcct aaaatggcgc ctgctatcct 1321 gagacgcccg acatcacctg tgtctagaga atgcaatagt agtacggata gctgtgactc 1381 cggtccttct aacacacctc ctgagataca cccggtggtc ccgctgtgcc ccattaaacc 1441 agttgccgtg agagttggtg ggcgtcgcca ggctgtggaa tgtatcgagg acttgcttaa 1501 cgagcctggg caacctttgg acttgagctg taaacgcccc aggccataag gtgtaaacct 1561 gtgattgcgt gtgtggttaa cgcctttgtt tgctgaatga gttgatgtaa gtttaataaa 1621 gggtgagata atgtttaact tgcatggcgt gttaaatggg gcggggctta aagggtatat 1681 aatgcgccgt gggctaatct tggttacatc tgacctcatg gaggcttggg agtgtttgga 1741 agatttttct gctgtgcgta acttgctgga acagagctct aacagtacct cttggttttg 1801 gaggtttctg tggggctcat cccaggcaaa gttagtctgc agaattaagg aggattacaa 1861 gtgggaattt gaagagcttt tgaaatcctg tggtgagctg tttgattctt tgaatctggg 1921 tcaccaggcg cttttccaag agaaggtcat caagactttg gatttttcca caccggggcg 1981 cgctgcggct gctgttgctt ttttgagttt tataaaggat aaatggagcg aagaaaccca 2041 tctgagcggg gggtacctgc tggattttct ggccatgcat ctgtggagag cggttgtgag 2101 acacaagaat cgcctgctac tgttgtcttc cgtccgcccg gcgataatac cgacggagga 2161 gcagcagcag cagcaggagg aagccaggcg gcggcggcag gagcagagcc catggaaccc 2221 gagagccggc ctggaccctc gggaatgaat gttgtacagg tggctgaact gtatccagaa 2281 ctgagacgca ttttgacaat tacagaggat gggcaggggc taaagggggt aaagagggag 2341 cggggggctt gtgaggctac agaggaggct aggaatctag cttttagctt aatgaccaga 2401 caccgtcctg agtgtattac ttttcaacag atcaaggata attgcgctaa tgagcttgat 2461 ctgctggcgc agaagtattc catagagcag ctgaccactt actggctgca gccaggggat 2521 gattttgagg aggctattag ggtatatgca aaggtggcac ttaggccaga ttgcaagtac 2581 aagatcagca aacttgtaaa tatcaggaat tgttgctaca tttctgggaa cggggccgag 2641 gtggagatag atacggagga tagggtggcc tttagatgta gcatgataaa tatgtggccg 2701 ggggtgcttg gcatggacgg ggtggttatt atgaatgtaa ggtttactgg ccccaatttt 2761 agcggtacgg ttttcctggc caataccaac cttatcctac acggtgtaag cttctatggg 2821 tttaacaata cctgtgtgga agcctggacc gatgtaaggg ttcggggctg tgccttttac 2881 tgctgctgga agggggtggt gtgtcgcccc aaaagcaggg cttcaattaa gaaatgcctc 2941 tttgaaaggt gtaccttggg tatcctgtct gagggtaact ccagggtgcg ccacaatgtg 3001 gcctccgact gtggttgctt catgctagtg aaaagcgtgg ctgtgattaa gcataacatg 3061 gtatgtggca actgcgagga cagggcctct cagatgctga cctgctcgga cggcaactgt 3121 cacctgctga agaccattca cgtagccagc cactctcgca aggcctggcc agtgtttgag 3181 cataacatac tgacccgctg ttccttgcat ttgggtaaca ggaggggggt gttcctacct 3241 taccaatgca atttgagtca cactaagata ttgcttgagc ccgagagcat gtccaaggtg 3301 aacctgaacg gggtgtttga catgaccatg aagatctgga aggtgctgag gtacgatgag 3361 acccgcacca ggtgcagacc ctgcgagtgt ggcggtaaac atattaggaa ccagcctgtg 3421 atgctggatg tgaccgagga gctgaggccc gatcacttgg tgctggcctg cacccgcgct 3481 gagtttggct ctagcgatga agatacagat tgaagtactg aaatgtgtgg gcgtggctta 3541 agggtgggaa agaatatata aggtgggggt cttatgtagt tttgtatctg ttttgcagca 3601 gccgccgccg ccatgagcac caactcgttt gatggaagca ttgtgagctc atatttgaca 3661 acgcgcatgc ccccatgggc cggggtgcgt cagaatgtga tgggctccag cattgatggt 3721 cgccccgtcc tgcccgcaaa ctctactacc ttgacctacg agaccgtgtc tggaacgccg 3781 ttggagactg cagcctccgc cgccgcttca gccgctgcag ccaccgcccg cgggattgtg 3841 actgactttg ctttcctgag cccgcttgca agcagtgcag cttcccgttc atccgcccgc 3901 gatgacaagt tgacggctct tttggcacaa ttggattctt tgacccggga acttaatgtc 3961 gtttctcagc agctgttgga tctgcgccag caggtttctg ccctgaaggc ttcctcccct 4021 cccaatgcgg tttaacggtc cgggccagag tggccgagca aaaggccagc aaaaggccag 4081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 4141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 4201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 4261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 4321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 4381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 4441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 4501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 4561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 4621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 4681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcggcca 4801 agaaggccag agtggcagag actgcattcg aaaacgtttg aattgataat tattatcatt 4861 tgcgggtcaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat 4921 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac 4981 cgaggcagtt ccataggatt gcaagatcct ggtatcggtc tgcgattccg actcgtccaa 5041 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac 5101 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt 5161 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat 5221 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac 5281 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac 5341 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga 5401 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt 5461 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc 5521 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac 5581 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg 5641 aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg 5701 tattactgtt tatgtaagca gacagtttta ttgttcatgc gaaaacgttt gaattgataa 5761 ttattatcat ttgcgggtcc tttccggcga tccgccttgt tacggggcgg cgacctcgcg 5821 ggttttcgct atttatgaaa attttccggt ttaaggcgtt tccgttcttc ttcgtcataa 5881 cttaatgttt ttatttaaaa taccctctga aaagaaagga aacgacagct gaaagcgagc 5941 tttttggcct ctgtcgtttc ctttctctgt ttttgtccgt ggaatgaaca atggaagtcg 6001 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagcgat 6061 atttaaatta a //