LOCUS Exported 9105 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express SaCas9 in bacteria with a 6xHis tag for purification. ACCESSION . VERSION . KEYWORDS p6XHis_NLS-SaCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9105) AUTHORS Soares Medeiros LC, South L, Peng D, Bustamante JM, Wang W, Bunkofske M, Perumal N, Sanchez-Valdez F, Tarleton RL TITLE Rapid, Selection-Free, High-Efficiency Genome Editing in Protozoan Parasites Using CRISPR-Cas9 Ribonucleoproteins. JOURNAL MBio. 2017 Nov 7;8(6). pii: e01788-17. doi: 10.1128/mBio.01788-17. PUBMED 29114029 REFERENCE 2 (bases 1 to 9105) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9105 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 28..483 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(115..134) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 325..346 /label=F1ori-F /note="F1 origin, forward primer" promoter 509..613 /gene="bla" /label=AmpR promoter CDS 614..1474 /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 complement(832..851) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1645..2233 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2134..2153 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2387..2404 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2419..2558 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2484..2503) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2644..2666 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2660..2851) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3660..4742) /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 4704..4723 /label=LacI-R /note="LacI, reverse primer" promoter complement(4743..4820) /gene="lacI" /label=lacI promoter primer_bind 5026..5045 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 5133..5152 /label=T7 /note="T7 promoter, forward primer" promoter 5133..5151 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5152..5176 /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)." RBS 5191..5213 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 5221..5547 /codon_start=1 /gene="trxA" /product="E. coli thioredoxin" /label=TrxA /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL DANLA" CDS 5569..5586 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 5596..5613 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 5620..5664 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 5680..5694 /codon_start=1 /product="enterokinase recognition and cleavage site" /label=enterokinase site /translation="DDDDK" CDS 5698..5718 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5728..8889 /codon_start=1 /product="Cas9 endonuclease from the Staphylococcus aureus Type II CRISPR/Cas system" /label=SaCas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ IIKKG" CDS 8965..8982 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(9035..9053) /label=T7 Term /note="T7 terminator, reverse primer" terminator 9049..9096 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg 61 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 121 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 181 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 241 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 301 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 361 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 421 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 481 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 1141 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact 1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 1441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 1501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 1561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 1621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 1681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 1741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 1801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 1861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 1921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 1981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 2041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 2101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 2161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 2221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 2281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 2341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 2401 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 2461 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat 2521 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg 2581 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 2641 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc 2701 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg 2761 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca 2821 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt 2881 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg 2941 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc 3001 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg 3061 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg 3121 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg 3181 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt 3241 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca 3301 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac 3361 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg 3421 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc 3481 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg 3541 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg 3601 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct 3661 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 3721 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag 3781 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc 3841 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa 3901 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc 3961 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc 4021 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca 4081 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc 4141 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt 4201 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag 4261 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta 4321 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc 4381 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt 4441 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc 4501 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac 4561 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc 4621 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg 4681 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc 4741 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt 4801 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag 4861 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg 4921 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa 4981 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat 5041 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta 5101 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg 5161 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat 5221 atgagcgata aaattattca cctgactgac gacagttttg acacggatgt actcaaagcg 5281 gacggggcga tcctcgtcga tttctgggca gagtggtgcg gtccgtgcaa aatgatcgcc 5341 ccgattctgg atgaaatcgc tgacgaatat cagggcaaac tgaccgttgc aaaactgaac 5401 atcgatcaaa accctggcac tgcgccgaaa tatggcatcc gtggtatccc gactctgctg 5461 ctgttcaaaa acggtgaagt ggcggcaacc aaagtgggtg cactgtctaa aggtcagttg 5521 aaagagttcc tcgacgctaa cctggccggt tctggttctg gccatatgca ccatcatcat 5581 catcattctt ctggtctggt gccacgcggt tctggtatga aagaaaccgc tgctgctaaa 5641 ttcgaacgcc agcacatgga cagcccagat ctgggtaccg atgacgacga caagatgccg 5701 aagaaaaagc gcaaggtcga agcgtccatg aaaaggaact acattctggg gctggacatc 5761 gggattacaa gcgtggggta tgggattatt gactatgaaa caagggacgt gatcgacgca 5821 ggcgtcagac tgttcaagga ggccaacgtg gaaaacaatg agggacggag aagcaagagg 5881 ggagccaggc gcctgaaacg acggagaagg cacagaatcc agagggtgaa gaaactgctg 5941 ttcgattaca acctgctgac cgaccattct gagctgagtg gaattaatcc ttatgaagcc 6001 agggtgaaag gcctgagtca gaagctgtca gaggaagagt tttccgcagc tctgctgcac 6061 ctggctaagc gccgaggagt gcataacgtc aatgaggtgg aagaggacac cggcaacgag 6121 ctgtctacaa aggaacagat ctcacgcaat agcaaagctc tggaagagaa gtatgtcgca 6181 gagctacagc tggaacggct gaagaaagat ggcgaggtga gagggtcaat taataggttc 6241 aagacaagcg actacgtcaa agaagccaag cagctgctga aagtgcagaa ggcttaccac 6301 cagctggatc agagcttcat cgatacttat atcgacctgc tggagactcg gagaacctac 6361 tatgagggac caggagaagg gagccccttc ggatggaaag acatcaagga atggtacgag 6421 atgctgatgg gacattgcac ctattttcca gaagagctga gaagcgtcaa gtacgcttat 6481 aacgcagatc tgtacaacgc cctgaatgac ctgaacaacc tggtcatcac cagggatgaa 6541 aacgagaaac tggaatacta tgagaagttc cagatcatcg aaaacgtgtt taagcagaag 6601 aaaaagccta cactgaaaca gattgctaag gagatcctgg tcaacgaaga ggacatcaag 6661 ggctaccggg tgacaagcac tggaaaacca gagttcacca atctgaaagt gtatcacgat 6721 attaaggaca tcacagcacg gaaagaaatc attgagaacg ccgaactgct ggatcagatt 6781 gctaagatcc tgactatcta ccagagttcc gaggacatcc aggaagagct gactaacctg 6841 aacagcgagc tgacccagga agagatcgaa cagattagta atctgaaggg gtacaccgga 6901 acacacaacc tgtccctgaa agctatcaat ctgattctgg atgagctgtg gcatacaaac 6961 gacaatcaga ttgcaatctt taaccggctg aagctggtac caaaaaaggt ggacctgagt 7021 cagcagaaag agatcccaac cacactggtg gacgatttca ttctgtcacc cgtggtcaag 7081 cggagcttca tccagagcat caaagtgatc aacgccatca tcaagaagta cggcctgccc 7141 aatgatatca ttatcgagct ggctagggag aagaacagca aggacgcaca gaagatgatc 7201 aatgagatgc agaaacgaaa ccggcagacc aatgaacgca ttgaagagat tatccgaact 7261 accgggaaag agaacgcaaa gtacctgatt gaaaaaatca agctgcacga tatgcaggag 7321 ggaaagtgtc tgtattctct ggaggccatc cccctggagg acctgctgaa caatccattc 7381 aactacgagg tcgatcatat tatccccaga agcgtgtcct tcgacaattc ctttaacaac 7441 aaggtgctgg tcaagcagga agagaactct aaaaagggca ataggactcc tttccagtac 7501 ctgtctagtt cagattccaa gatctcttac gaaaccttta aaaagcacat tctgaatctg 7561 gccaaaggaa agggccgcat cagcaagacc aaaaaggagt acctgctgga agagcgggac 7621 atcaacagat tctccgtcca gaaggatttt attaaccgga atctggtgga cacaagatac 7681 gctactcgcg gcctgatgaa tctgctgcga tcctatttcc gggtgaacaa tctggatgtg 7741 aaagtcaagt ccatcaacgg cgggttcaca tcttttctga ggcgcaaatg gaagtttaaa 7801 aaggagcgca acaaagggta caagcaccat gccgaagatg ctctgattat cgcaaatgcc 7861 gacttcatct ttaaggagtg gaaaaagctg gacaaagcca agaaagtgat ggagaaccag 7921 atgttcgaag agaagcaggc cgaatctatg cccgaaatcg agacagaaca ggagtacaag 7981 gagattttca tcactcctca ccagatcaag catatcaagg atttcaagga ctacaagtac 8041 tctcaccggg tggataaaaa gcccaacaga gagctgatca atgacaccct gtatagtaca 8101 agaaaagacg ataaggggaa taccctgatt gtgaacaatc tgaacggact gtacgacaaa 8161 gataatgaca agctgaaaaa gctgatcaac aaaagtcccg agaagctgct gatgtaccac 8221 catgatcctc agacatatca gaaactgaag ctgattatgg agcagtacgg cgacgagaag 8281 aacccactgt ataagtacta tgaagagact gggaactacc tgaccaagta tagcaaaaag 8341 gataatggcc ccgtgatcaa gaagatcaag tactatggga acaagctgaa tgcccatctg 8401 gacatcacag acgattaccc taacagtcgc aacaaggtgg tcaagctgtc actgaagcca 8461 tacagattcg atgtctatct ggacaacggc gtgtataaat ttgtgactgt caagaatctg 8521 gatgtcatca aaaaggagaa ctactatgaa gtgaatagca agtgctacga agaggctaaa 8581 aagctgaaaa agattagcaa ccaggcagag ttcatcgcct ccttttacaa caacgacctg 8641 attaagatca atggcgaact gtatagggtc atcggggtga acaatgatct gctgaaccgc 8701 attgaagtga atatgattga catcacttac cgagagtatc tggaaaacat gaatgataag 8761 cgcccccctc gaattatcaa aacaatcgcc tctaagactc agagtatcaa aaagtactca 8821 accgacattc tgggaaacct gtatgaggtg aagagcaaaa agcaccctca gattatcaaa 8881 aagggctaac cgggcttctc ctcaaccatg gcgatatcgg atccgaattc gagctccgtc 8941 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac 9001 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc 9061 cttggggcct ctaaacgggt cttgaggggt tttttgctga aagga //