LOCUS Exported 9579 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tetracycline inducible expression of guide RNA targeted to AAVS1 locus. ACCESSION . VERSION . KEYWORDS pAAV-Puro_siKO SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9579) AUTHORS Bertero A, Pawlowski M, Ortmann D, Snijders K, Yiangou L, Cardoso de Brito M, Brown S, Bernard WG, Cooper JD, Giacomelli E, Gambardella L, Hannan NR, Iyer D, Sampaziotis F, Serrano F, Zonneveld MC, Sinha S, Kotter M, Vallier L TITLE Optimized inducible shRNA and CRISPR/Cas9 platforms for in vitro studies of human development using hPSCs. JOURNAL Development. 2016 Dec 1;143(23):4405-4418. PUBMED 27899508 REFERENCE 2 (bases 1 to 9579) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9579 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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 186..208 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 205..221 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 242..260 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" misc_feature 295..1098 /label=HA-L /note="left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" misc_feature 1105..1130 /label=SA /note="splice acceptor site" CDS 1154..1207 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 1217..1816 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1217..1236) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1713..1733 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 1854..2078 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 2222..2241 /label=H1 /note="Human H1 promoter, forward primer" protein_bind 2287..2305 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" misc_RNA 2340..2415 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 2494..2797 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2799..3076 /label=chicken beta-actin promoter intron 3078..4095 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 4019..4041 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 4103..4122 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 4166..4186 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 4840..4951 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" misc_feature 5081..5917 /label=HA-R /note="right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" primer_bind complement(5951..5970) /label=T7 /note="T7 promoter, forward primer" promoter complement(5952..5970) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5977..5994) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5977..5993) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5986..6008) /label=M13/pUC Forward /note="In lacZ gene" CDS 6131..6433 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 6312..6331 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 6782..7576 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6836..6855) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7446..7465 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS complement(7826..8686) /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 8449..8468 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8810..9398 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9299..9318 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9552..9569 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca 241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt 301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg 361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct 421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc 481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg 541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc 601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg 661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt 721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt 781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg 841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca 901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat 961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc 1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc 1081 tccaccccac agtggggcaa gcttctgacc tcttctcttc ctcccacagg gcctcgagag 1141 ttctggcagc ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc 1201 cggccctagg ctcgagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 1261 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 1321 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 1381 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 1441 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 1501 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 1561 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 1621 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 1681 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 1741 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 1801 caagcccggt gcctgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 1861 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1921 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1981 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 2041 caatagcagg catgctgggg atgcggtggg ctctatgggt caattcgaac gctgacgtca 2101 tcaacccgct ccaaggaatc gcgggcccag tgtcactagg cgggaacacc cagcgcgcgt 2161 gcgccctggc aggaagatgg ctgtgaggga caggggagtg gcgccctgca atatttgcat 2221 gtcgctatgt gttctgggaa atcaccataa acgtgaaatg tctttggatt tgggaatctt 2281 ataagttccc tatcagtgat agagatccct ggtgcaggtg gactgactca cctgcacctg 2341 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg 2401 gcaccgagtc ggtgcttttt tgtcgacctc gactagttat taatagtaat caattacggg 2461 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 2521 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 2581 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 2641 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga 2701 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg 2761 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg 2821 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta 2881 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg 2941 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg 3001 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg 3061 aagcgcgcgg cgggcgggga gtcgctgcga cgctgccttc gccccgtgcc ccgctccgcc 3121 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg 3181 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct 3241 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc 3301 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg 3361 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc 3421 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa 3481 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg 3541 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg 3601 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggctggtg 3661 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc 3721 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg 3781 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct 3841 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg 3901 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc 3961 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg 4021 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc 4081 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc 4141 aaagaattaa ttcggatcca ccatgcccaa gaaaaagcgg aaggtgtccc ggctggacaa 4201 gagcaaagtg atcaacagcg ccctggaact gctgaacgaa gtgggcatcg agggcctgac 4261 cacccggaag ctggcccaga aactgggcgt ggaacagccc accctgtact ggcacgtgaa 4321 gaacaagcgg gccctgctgg acgccctggc catcgagatg ctggaccggc accacacaca 4381 cttttgcccc ctggaaggcg aaagctggca ggacttcctg cggaacaacg ccaagagctt 4441 cagatgcgcc ctgctgagcc accgggacgg cgccaaagtg cacctgggca ccagacccac 4501 cgagaagcag tacgagacac tggaaaacca gctggccttc ctgtgccagc agggcttcag 4561 cctggaaaac gccctgtacg ccctgagcgc cgtgggccac tttaccctgg gctgcgtgct 4621 ggaagatcag gaacaccagg tcgccaaaga ggaaagagag acacccacca ccgacagcat 4681 gccccccctg ctgagacagg ccatcgagct gttcgatcat caaggcgccg agcccgcctt 4741 cctgttcggc ctggaactga tcatctgcgg cctcgagaag cagctgaagt gcgagagcgg 4801 ctcccccaga tgaacgcgta gctcgctgat cagcctcgac tgtgccttct agttgccagc 4861 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg 4921 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc 4981 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg 5041 ctgggggtct cgaggggggg cccggacagt actaagcttt actagggaca ggattggtga 5101 cagaaaagcc ccatccttag gcctcctcct tcctagtctc ctgatattgg gtctaacccc 5161 cacctcctgt taggcagatt ccttatctgg tgacacaccc ccatttcctg gagccatctc 5221 tctccttgcc agaacctcta aggtttgctt acgatggagc cagagaggat cctgggaggg 5281 agagcttggc agggggtggg agggaagggg gggatgcgtg acctgcccgg ttctcagtgg 5341 ccaccctgcg ctaccctctc ccagaacctg agctgctctg acgcggctgt ctggtgcgtt 5401 tcactgatcc tggtgctgca gcttccttac acttcccaag aggagaagca gtttggaaaa 5461 acaaaatcag aataagttgg tcctgagttc taactttggc tcttcacctt tctagtcccc 5521 aatttatatt gttcctccgt gcgtcagttt tacctgtgag ataaggccag tagccagccc 5581 cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc ctttgtgaga 5641 atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct actccctttc tctttctcca 5701 tccttctttc cttaaagagt ccccagtgct atctgggaca tattcctccg cccagagcag 5761 ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt tgagtccttg gcaagcccag 5821 gagaggcgct caggcttccc tgtccccctt cctcgtccac catctcatgc ccctggctct 5881 cctgcccctt ccctacaggg gttcctggct ctgctctaag ggcgaattcg cggccgctaa 5941 attcaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg 6001 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 6061 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 6121 atacgtacgg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt 6181 ggatgtacag agtgatatta ttgacacgcc ggggcgacgg atggtgatcc ccctggccag 6241 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga 6301 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga 6361 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt 6421 ctggggaata taaatgtcag gcatgagatt atcaaaaagg atcttcacct agatcctttt 6481 cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc 6541 tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac 6601 atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg 6661 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc 6721 aaggatctga tggcgcaggg gatcaagctc tgatcaagag acaggatgag gatcgtttcg 6781 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 6841 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 6901 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 6961 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 7021 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 7081 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 7141 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 7201 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 7261 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga 7321 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 7381 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 7441 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 7501 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 7561 tgacgagttc ttctgaatta ttaacgctta caatttcctg atgcggtatt ttctccttac 7621 gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat gtgcgcggaa 7681 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agattatcaa 7741 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 7801 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 7861 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 7921 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 7981 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 8041 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 8101 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8161 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8221 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8281 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8341 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 8401 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 8461 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 8521 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 8581 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 8641 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 8701 aatattattg aagcatttat cagggttatt gtctcatgac caaaatccct taacgtgagt 8761 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 8821 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 8881 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 8941 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 9001 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 9061 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 9121 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 9181 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 9241 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 9301 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 9361 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 9421 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 9481 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 9541 cgaccgagcg cagcgagtca gtgagcgagg aagcggaag //