LOCUS Exported 9703 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR/Cas9 NICKASE plasmid against human NFATc2 (1/2). ACCESSION . VERSION . KEYWORDS NFATc2-CRISPR-Nick 1/2 (PX461-EF1a-pSpCas9n(BB)-2A-GFP) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9703) AUTHORS Gabriel CH, Gross F, Karl M, Stephanowitz H, Hennig AF, Weber M, Gryzik S, Bachmann I, Hecklau K, Wienands J, Schuchhardt J, Herzel H, Radbruch A, Krause E, Baumgrass R TITLE Identification of Novel Nuclear Factor of Activated T Cells (NFAT)-Associated Proteins in T cells. JOURNAL J Biol Chem. 2016 Sep 16. pii: jbc.M116.739326. PUBMED 27637333 REFERENCE 2 (bases 1 to 9703) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9703 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 272..347 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 460..1638 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 691..1629 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1586..1606 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 1659..1668 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1668..1733 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1740..1760 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1785..5885 /codon_start=1 /product="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9(D10A) /note="generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5886..5933 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 5949..6002 /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 6003..6716 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 6003..6716 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(6045..6066) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6306..6325) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6653..6674 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(6744..6761) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6750..6957 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6966..7106 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 6966..7095 /label=AAV2 ITR rep_origin 7181..7636 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7268..7287) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7478..7499 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7653..7672) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7772..7794 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7832..7850) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7918..8022 /gene="bla" /label=AmpR promoter CDS 8023..8883 /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(8241..8260) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9054..9642 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9543..9562 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg ggccgcagaa gtttctgagc ggttttagag ctagaaatag caagttaaaa 301 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag 361 agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc 421 agacaaatgg ctctagtggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg 481 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc 541 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc 601 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt 661 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct 721 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac 781 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct 841 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc 901 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt 961 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg 1021 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt 1081 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga 1141 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc 1201 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg 1261 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg 1321 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt 1381 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt 1441 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg 1501 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt 1561 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt 1621 ccatttcagg tgtcgtgagg aatttcgaca tttccggtgc caccatggac tataaggacc 1681 acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc 1741 caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca 1801 tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg 1861 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc 1921 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa 1981 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca 2041 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg 2101 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg 2161 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg 2221 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc 2281 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc 2341 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg 2401 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg 2461 cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg 2521 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga 2581 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg 2641 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga 2701 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg 2761 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt 2821 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag 2881 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg 2941 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca 3001 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg 3061 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc 3121 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca 3181 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt 3241 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg 3301 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag 3361 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg 3421 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg 3481 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt 3541 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc 3601 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg 3661 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag 3721 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga 3781 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg 3841 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg 3901 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc 3961 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc 4021 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga 4081 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg 4141 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc 4201 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg 4261 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc 4321 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga 4381 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc 4441 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg 4501 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga 4561 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg 4621 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc 4681 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg 4741 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg 4801 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc 4861 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca 4921 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg 4981 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc 5041 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca 5101 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg 5161 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg 5221 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca 5281 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct 5341 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg 5401 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg 5461 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg 5521 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg 5581 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc 5641 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg 5701 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact 5761 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc 5821 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag 5881 gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg 5941 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc 6001 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 6061 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 6121 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 6181 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 6241 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 6301 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 6361 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 6421 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 6481 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 6541 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 6601 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 6661 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggaat 6721 tctaactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt 6781 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 6841 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 6901 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg 6961 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac 7021 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag 7081 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct 7141 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca 7201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 7261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 7321 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 7381 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt 7441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 7501 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg 7561 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 7621 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 7681 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 7741 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 7801 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt 7861 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 7921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 7981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 8041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 8101 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 8161 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 8221 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 8281 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 8341 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 8401 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 8461 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 8521 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 8581 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 8641 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 8701 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca 8761 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 8821 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 8881 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 8941 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 9001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 9061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 9121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 9181 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac 9241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 9301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 9361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 9421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 9481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 9541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 9601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 9661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt //