LOCUS Exported 10706 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression plasmid for protein purification. ACCESSION . VERSION . KEYWORDS 6His-MBP-TEV-huAsCpf1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10706) AUTHORS Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS, Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F TITLE Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. JOURNAL Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5. PUBMED 27918548 REFERENCE 2 (bases 1 to 10706) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10706 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1334..1525 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind complement(1519..1541) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 1627..1766 /label=bom /note="basis of mobility region from pBR322" primer_bind 1682..1701 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(1781..1798) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1952..2540) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2032..2051) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2711..3571) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3334..3353 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3572..3676) /gene="bla" /label=AmpR promoter rep_origin complement(3702..4157) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3839..3860) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4051..4070 /label=F1ori-R /note="F1 origin, reverse primer" terminator 4194..4241 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 4237..4255 /label=T7 Term /note="T7 terminator, reverse primer" CDS complement(4308..4325) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS complement(4341..4385) /codon_start=1 /product="peptide tag that allows for enzymatic biotinylation" /label=AviTag(TM) /translation="GLNDIFEAQKIEWHE" CDS complement(4386..4409) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS complement(4434..4514) /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" CDS complement(4521..4568) /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS complement(4569..8489) /codon_start=1 /product="CRISPR-associated protein from Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)" /label=AsCpf1 /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES KDLKLQNGISNQDWLAYIQELRN" regulatory 4840..4849 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(8490..8510) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS complement(8517..8534) /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" regulatory 8687..8696 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(9651..9668) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" RBS 9682..9704 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 9719..9743 /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 complement(9743..9762) /label=T7 /note="T7 promoter, forward primer" promoter complement(9744..9762) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(9846..9865) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 10071..10148 /gene="lacI" /label=lacI promoter CDS 10149..525 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(10168..10187) /label=LacI-R /note="LacI, reverse primer" ORIGIN 1 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact 61 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt 121 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac 181 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg 241 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta 301 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa 361 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga 421 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 541 tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa 601 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt 661 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga 721 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt 781 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca 841 ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag 901 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga 961 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt 1021 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat 1081 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt 1141 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc 1201 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat 1261 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta 1321 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag 1381 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga 1441 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt 1501 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct 1561 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg 1621 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat 1681 gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat 1741 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc 1801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 1861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 1921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 1981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 2041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 2101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 2161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 2221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 2281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 2341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 2401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 2461 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 2521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 2581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 2641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 2701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 2761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 2821 catctggccc cagtgctgca atgataccgc gactaccacg ctcaccggct ccagatttat 2881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 2941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 3001 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta 3061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 3121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 3181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 3241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 3301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 3361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 3421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 3481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 3541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 3601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 3661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat 3721 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 3781 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 3841 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 3901 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 3961 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 4021 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 4081 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 4141 gccgctacag ggcgcgtccc attcgccaat ccggatatag ttcctccttt cagcaaaaaa 4201 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca 4261 gcagccaact cagcttcctt tcgggctttg ttagcagccg gatctcagtg gtggtggtgg 4321 tggtgctcga caagctttta ctcgtgccat tcaattttct gggcttcaaa gatatcgttc 4381 aggcctttat catcatcatc tttataatcg ctaccaccac cctcgaggct ttaggcatag 4441 tcggggacat catatgggta tgcataatca ggcacgtcgt agggataagc gtaatctgga 4501 acatcgtatg ggtaggatcc ctttttcttt tttgcctggc cggccttttt cgtggccgcc 4561 ggccttttgt tgcgcagctc ctggatgtag gccagccagt cctgattgga gatgccgttc 4621 tgcagcttca gatccttgct ctccttcagg tgattcagca gcagctggcc cttcagggcg 4681 atgtggtagg cgccattggc atcggcgtcc atgggccact ctgggttctg aaaccgggag 4741 tcgaagcaca cgccattcag atcgcgcacg gggctgttga tatagtcctc gcctgtggcg 4801 gcattggagt tccgcatctg cagcacgctg cggatcaggg ccaccatggt gtcgatggcg 4861 tgagaatcgt cattctccag cagctttggc aggatgttgg agccatccct gaacacgatg 4921 cccttctcct ccagcagggc gatcagctcg ttggcaggat acaggtcccg gtatctgccg 4981 gtgaatctgt gattctcgat cactggcacg attctcttgc cggcgatgaa aggggtgccc 5041 ttggcgtcaa actgtgtctc gttcttctcg aacacgatat cccatgcagg cataaagccg 5101 ggcaggcccc tctggaagga cagatttctg ttcatcttaa agtgcaggat gaagtcgccg 5161 gttttcacgt cgtagtgcag aaagtcgaag ccctccagga agtgcttgcg gctctcgtga 5221 ttcttgatgg ttttccacac gaaggggtcc acgaagccgg tcaggggatc gatcttagat 5281 gtatatgggg caggcacgta aaacaggaag ccagactggg tgcccatctt ggcaaaggag 5341 gtgaactggt ctgtcagctg gtatgggttc agcacgcctc ccactttctc tgctggatag 5401 tccttcagca ccaggcaatt cagcttatcg atcagcatct tctcgaactg ctggtacacg 5461 gccttctcgg cgatgccggt cctcttgctc ttaaagccga aattcaggtt ctccagcacc 5521 accacggcct ggtagtggat catcaggtcc acgatctcgt ggatgacctg gctcagatag 5581 ccctgcttca gatccttgat tgtgcccacc acagaccagg cctgccttgc tgccaccctc 5641 tccttctccc tgttgtccag cttcttctgg taatcaaact gctggatggt gttcaggctc 5701 cgctgctcca ggatcttgcc ggtggagtcg atcactgtga tatagatcag gtttctctcg 5761 ccccgatcga tgccgatgat aggtgtctcg gggtgctcct tcaggtaggc attcaccctc 5821 tggttgaact tagatgggga attggcggcc tgatagttca gtgtgatagg cacgtggaaa 5881 aagaacttgt cgctggtaaa gcgcctatcc ttgatgatct cgtgagacac ctccttggtg 5941 atcacgttgg gcagcagggc cctggcctca tcagacaggt cgtgggacag tctgtgattc 6001 acatagtcgt acagctcctg gtacagggtg tcggggattg gggttttctg atccttcagc 6061 ttcttgttca gcatcttctc tcccagccgg tgtgccatcc tcttcatcct ggacttaggg 6121 cggtagaaca gctcggcctg gccattcagc ttgatgcttg tcttggccag gttctctgga 6181 gaaaacaggc cggtccaata cagtgtgtgc agattaggct tgccgtggtg gcccttggca 6241 aagtccttgt tatagatctg gaacaggtac agcttgcctg tctccacggc atccatgatc 6301 tccttctcgg cgattctctg gaagctgatg tggtacagca ggggattcag ctcggcatag 6361 tactcgccca ggtccttata ctgagaggat ggccgcaggc tagacagatc gatagaggtt 6421 gtcttggtat acttggacag aaaatccctt gtgaagtcga tccacttgca cagggcctct 6481 ctgtagccct tctggtcgcc ggttttcttg gcgtaggctg tctgaaactt ctttggctcc 6541 ttctcaggat tgttcaggtc gtagatctcc tttgtgatct ccagaggctc gatgaaattg 6601 ttggacagca ggatgggggt tgtgtgggtc tgaaagtggg ctgtcacggc cttcagctgg 6661 gtgctgcact ttgggatcat cttggcggca tcagggaagt agtcatagta catcttatca 6721 aagccctcgc tggttttctc tgtgggctcg aagctcaggg ccttatacct gcccttctgc 6781 tttggcatga tgcccagata gtacaggccg ttcttcacaa acaggatggc gccattgttc 6841 ttctccttat tcacgtccca gccagaggcc agtgtaggca tctgaaagtt cagcttgaac 6901 ttctccacgg agtagggctt cttggtggca taatttctgg ccttgttgta gaagctcaga 6961 gaaggctcca tctccagctt gatgccggtc agccgggcag agaactcggg gtccacctcg 7021 ttggactcat ccacggcaaa ccagtccagc aggtggtaca ggcccagcag gctgtccagc 7081 tgagacttca ggatctcctt ctcctcctgc ttcttcaggg ttgtaggcag tggctgatcc 7141 agggcggcgt gtgcgtggga caggatctcg ctggttttct gcttgaaggc ctcgctcagc 7201 tccttgcctg cggcagagat gatctcctgc aggttgatat cctcgtgctt caggctgcgc 7261 tgcaccttct ccttggcaga cttggtgatc ttgcctgtca gctcggagat tctccgctca 7321 tacagggcat tcctcagtgt atcccagtgg tcgcacaggg cgctgctgat tgtctccagc 7381 ttcttgtggc tgatgaagat gtgtgtcagg tcgatgctgt tcagctcgtt aaacagggcc 7441 tcggctgtct ccagcacgtt ctcgtttctc agcagtgtct tgtacttgca gaaggactgg 7501 atcacttcct cgtcgctctt aaactcctcc aggatgaaag acagggtgtt cctatcggac 7561 aggatctgct taaacagggg gatgaatctg tgtggcaggg aggcgatgat gtgggctgtc 7621 tcatcattct tctggatggc cagattcagc acctcgttca ggcccttgat cttctcggtg 7681 cctgcctccc gagagattcc tcccagcagc tggttataca ggtcgatctg ggtctgtgtc 7741 agcagctggt tataaaaagg gaaggaaaac acctcctcga tggaggtgct cacgaagatg 7801 ccgatggcct tcttcacgtt ctcaaagtgc tcccgcaggc tgggcacggc ggtgatcagg 7861 cgtgtgaaga tgtgacaatt ctccttaaac ttggggaagt tgtcctgcac gatgcggtgt 7921 gggatggctg tgctgatatc ctcggcgctg aacacgttct tcctgttctc ataaaagccg 7981 gagaagtagg ttgtaaactt gtcgaagctc cgcagcaggg cgttctcgtg ctcggttgtg 8041 gtcacggtgc ccagctgctt cagcaccttg ccattaaaca gctcggcctt gaacaggccc 8101 ttgtagatct cggcgtgtct cttattgatg gcatcggtca ggttgtctgt ccggccgatg 8161 aagtagtcgt ggatggcatt gcgatatgtg gcctgctcct cgatcagggc gttccttgtc 8221 tcctcggttt tctcctttct ataggagtcg atggcggcgc tcaggttctc ccaatccagc 8281 tgcaccagct gcaggcactg gtcggcatag gtcttgtaga tccgatcgat gatgggcttc 8341 agctccttgt agtgatcatt gcgggccttg tcctcctcga tgaagccctg ctcctggatg 8401 tgcttcaggg tcttgccctg tgggatcagc tcaaaccgca gtgtcttgct cacctgatac 8461 aggttggtaa agccctcgaa ctgtgtcatt ccctgaaaat acagattttc gctagcgcta 8521 ccacgcggaa ccagggatcc ggtgcttgca ccggtctggg catctttcag ggcttcatca 8581 acggtctgac gacctgatgc ggcattaatc actgcggtac gaacggcata ccaaaatgcg 8641 ctcatctgcg gaatattcgg cataatttcg cctttctgtg cattttccat ggtggctgca 8701 atacgcggat ctttaaccag ttcttcttca tagcttttca gtgcaactgc acccagcggt 8761 ttatctttat tcactgcttc cagaccttca tcggtcagca gatagttttc cagaaattct 8821 tttgccagtt ctttgttcgg gcttgctgca ttaatacctg cgctcagaac accaacaaac 8881 ggtttgctcg gctgaccttt aaatgtcggc agaacggtaa caccataatt cactttgctg 8941 gtatcaatat ttgaccatgc ccacggacca ttaattgtca ttgcggtttc acctttgtta 9001 aatgctgctt ctgcaatgct ataatcggta tcggcgttca tgtgtttgtt tttaatcaga 9061 tcaaccagaa aggtcagacc ggcttttgca cccgcattat caacacccac gtctttgata 9121 tcatatttgc cgttttcata tttgaatgca tagccaccat ctgctgcaat cagaggccag 9181 gtaaaatacg gttcttgcag attaaacatc agtgcgcttt taccttttgc tttcagttct 9241 ttatccagtg ccggaatttc ttcccaggtt ttaggcggat tcggcagcag atctttgtta 9301 tagatcaggc tcagtgcttc aactgcaatc ggatatgcaa tcagtttacc gttatagcga 9361 actgcatccc aggtaaacgg atacagtttg tcctgaaatg ctttatccgg tgtaatttct 9421 gccagcagac cgctctgtgc ataaccacca aaacgatcat gtgcccaaaa gataatatcc 9481 ggaccatcac cggttgctgc aacctgcgga aatttttctt ccagtttatc cggatgttca 9541 acggtcactt taatgccggt atctttttcg aattttttgc caacttctgc cagaccatta 9601 tagcctttat cgccattgat ccaaataacc agtttgcctt cttcgatttt gtgatggtga 9661 tgatgatgac ccatatgtat atctccttct taaagttaaa caaaattatt tctagagggg 9721 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat 9781 ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc 9841 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat 9901 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc 9961 catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact 10021 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg 10081 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg 10141 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc 10201 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag 10261 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg 10321 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc 10381 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga 10441 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac 10501 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag 10561 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca 10621 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg 10681 gtcaccagca aatcgcgctg ttagcg //