LOCUS Exported 9522 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentivirus production of CheRiff channelrhodopsin actuator. ACCESSION . VERSION . KEYWORDS pMOS010: pLX304-CheRiff-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9522) AUTHORS Hochbaum DR, Zhao Y, Farhi SL, Klapoetke N, Werley CA, Kapoor V, Zou P, Kralj JM, Maclaurin D, Smedemark-Margulies N, Saulnier JL, Boulting GL, Straub C, Cho YK, Melkonian M, Wong GK, Harrison DJ, Murthy VN, Sabatini BL, Boyden ES, Campbell RE, Cohen AE TITLE All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins. JOURNAL Nat Methods. 2014 Aug;11(8):825-33. doi: 10.1038/nmeth.3000. Epub 2014 Jun 22. PUBMED 24952910 REFERENCE 2 (bases 1 to 9522) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9522 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..181 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 228..353 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 846..1079 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1264..1308 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 1483..1983 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 1902..1920 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 1995..2390 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 2450..2567 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2684..2987 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2988..3191 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3142..3162 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 3214..3238 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 3270..3279 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3276..4223 /codon_start=1 /product="blue light-activated optogenetic actuator derived from Scherffelia dubia channelrhodopsin (Hochbaum et al., 2014)" /label=CheRiff /translation="MGGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQ CSSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKV ILEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALL VSDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLV LAMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKI HEHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV" CDS 4293..5006 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(4335..4356) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4596..4615) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4943..4964 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(5039..5063) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5065..5106 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 5148..5736 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5201..5221) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5619..5630) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5808..6041 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6119..6240 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6156..6175) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6210..6229 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6280..6415 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6342..6361 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6435..6454) /label=T7 /note="T7 promoter, forward primer" promoter complement(6436..6454) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6464..6481) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6464..6480) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6473..6495) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6622..7077 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6709..6728) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6919..6940 /label=F1ori-F /note="F1 origin, forward primer" promoter 7103..7207 /gene="bla" /label=AmpR promoter CDS 7208..8068 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(7426..7445) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8239..8827 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8728..8747 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8981..8998 /label=L4440 /note="L4440 vector, forward primer" protein_bind 9115..9136 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9151..9181 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9189..9205 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 9194..9216 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 9213..9229 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 9213..9229 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 9248..9268 /label=T3 /note="T3 promoter, forward primer" promoter 9250..9268 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 9296..9522 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" ORIGIN 1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca 241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc 301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta 361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa 421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc 481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc 541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg 601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc 661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga 721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccttgcgcct tttccaaggc 1501 agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc 1561 gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc 1621 gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc 1681 ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc 1741 gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat 1801 agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg 1861 gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag 1921 cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc 1981 cagggggtac caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag 2041 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc 2101 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt 2161 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta 2221 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg 2281 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga 2341 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc ctgcagctgc 2401 agtagtaaga attctagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa 2461 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca 2521 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca 2581 gggacagcag agatccactt tggcgccggc tcgagggggc ccggggttat taatagtaat 2641 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 2701 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 2761 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 2821 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 2881 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 2941 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 3001 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 3061 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 3121 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 3181 taagcagagc tctctggcta actgtcggga tcaacaagtt tgtacaaaaa agcaggctgg 3241 cgccggaacc aattcagtcg actggatccg ccaccatggg cggagctcct gctccagacg 3301 ctcacagcgc cccacctgga aacgattctg ccggaggcag tgagtaccat gccccagctg 3361 gatatcaagt gaatccaccc taccaccccg tgcatgggta tgaggaacag tgcagctcca 3421 tctacatcta ctatggggcc ctgtgggagc aggaaacagc taggggcttc cagtggtttg 3481 ccgtgttcct gtctgccctg tttctggctt tctacggctg gcacgcctat aaggccagcg 3541 tgggatggga ggaagtgtac gtgtgctccg tggagctgat caaagtgatt ctggagatct 3601 atttcgagtt caccagtcct gctatgctgt tcctgtacgg agggaacatt accccatggc 3661 tgagatatgc cgaatggctg ctgacatgtc ccgtgatcct gattcatctg tctaacatca 3721 ccggcctgag tgaggcatac aataagcgga caatggctct gctggtgtcc gacctgggaa 3781 ctatttgcat gggagtgaca gccgctctgg ccactgggtg ggtgaagtgg ctgttttact 3841 gtatcggcct ggtgtatgga acccagacat tctacaacgc tggaatcatc tacgtggagt 3901 cttactatat catgcctgcc ggcggctgta agaaactggt gctggccatg actgccgtgt 3961 actattctag ttggctgatg tttcccggcc tgttcatctt tgggcctgaa ggcatgcaca 4021 ccctgagcgt ggctgggtcc actattggcc ataccatcgc cgacctgctg tccaagaata 4081 tttggggact gctggggcac ttcctgcgga tcaaaattca cgagcatatc attatgtacg 4141 gcgatatcag gagaccagtg agctcccagt ttctgggacg caaggtggac gtgctggcct 4201 tcgtgacaga ggaagataaa gtggcggccg ccaagagcag gatcaccagc gagggcgagt 4261 acatccccct ggaccagatc gacatcaacg tggtgagcaa gggcgaggag ctgttcaccg 4321 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 4381 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atttgcacca 4441 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 4501 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 4561 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 4621 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 4681 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 4741 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 4801 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 4861 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 4921 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 4981 ctctcggcat ggacgagctg tacaagtaag aattcgcggc cgcactcgag atatctagac 5041 ccagctttct tgtacaaagt ggttggtaag cctatcccta accctctcct cggtctcgat 5101 tctacgtagt aatgagctag cgctaaccgg tggcgcgtta agtcgacaat caacctctgg 5161 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 5221 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 5281 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 5341 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 5401 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 5461 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 5521 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca 5581 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 5641 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 5701 agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta 5761 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 5821 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca 5881 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 5941 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6001 atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg 6061 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6121 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6181 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6241 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6301 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6361 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg 6421 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac 6481 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc 6541 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc 6601 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 6661 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 6721 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 6781 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg 6841 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg 6901 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct 6961 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg 7021 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg 7081 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 7141 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 7201 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 7261 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 7321 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 7381 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 7441 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 7501 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 7561 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 7621 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 7681 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 7741 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 7801 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 7861 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 7921 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 7981 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 8041 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 8101 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 8161 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 8221 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 8281 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 8341 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 8401 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 8461 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 8521 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 8581 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 8641 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 8701 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 8761 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 8821 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 8881 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 8941 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 9001 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg 9061 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt 9121 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt 9181 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc 9241 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag 9301 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta 9361 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc 9421 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat 9481 tgcagagata ttgtatttaa gtgcctagct cgatacataa ac //