LOCUS Exported 10549 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Protein expression plasmid for 2xNLS FnCpf1 in pET-21a backbone. ACCESSION . VERSION . KEYWORDS pET-21a_2xNLS_FnCpf1_MBP_protein_expression SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10549) AUTHORS Liu P, Luk K, Shin M, Idrizi F, Kwok S, Roscoe B, Mintzer E, Suresh S, Morrison K, Frazao JB, Bolukbasi MF, Ponnienselvan K, Luban J, Zhu LJ, Lawson ND, Wolfe SA TITLE Enhanced Cas12a editing in mammalian cells and zebrafish. JOURNAL Nucleic Acids Res. 2019 Mar 20. pii: 5403491. doi: 10.1093/nar/gkz184. PUBMED 30892626 REFERENCE 2 (bases 1 to 10549) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10549 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 552..571 /label=LacI-R /note="LacI, reverse primer" promoter complement(591..668) /gene="lacI" /label=lacI promoter primer_bind 874..893 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 977..996 /label=T7 /note="T7 promoter, forward primer" promoter 977..995 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 996..1020 /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 1035..1057 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4965..5012 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 5019..5099 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" CDS 5100..5120 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5130..5150 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" primer_bind 6224..6247 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 6257..6274 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6327..6345) /label=T7 Term /note="T7 terminator, reverse primer" terminator 6341..6388 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 6425..6880 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6512..6531) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6722..6743 /label=F1ori-F /note="F1 origin, forward primer" promoter 6906..7010 /gene="bla" /label=AmpR promoter CDS 7011..7871 /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(7229..7248) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8042..8630 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8531..8550 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8784..8801 /label=L4440 /note="L4440 vector, forward primer" misc_feature 8816..8955 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(8881..8900) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9041..9063 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(9057..9248) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(10057..590) /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" ORIGIN 1 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc 61 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg 121 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct 181 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt 241 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc 301 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc 361 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc 421 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact 481 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga 541 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc 601 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg 661 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag 721 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc 781 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat 841 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc 901 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat 961 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc 1021 cctctagaaa taattttgtt taactttaag aaggagatat acatatgagc atctaccagg 1081 agttcgtcaa caagtattca ctgagtaaga cactgcggtt cgagctgatc ccacagggca 1141 agacactgga gaacatcaag gcccgaggcc tgattctgga cgatgagaag cgggcaaaag 1201 actataagaa agccaagcag atcattgata aataccacca gttctttatc gaggaaattc 1261 tgagctccgt gtgcatcagt gaggatctgc tgcagaatta ctcagacgtg tacttcaagc 1321 tgaagaagag cgacgatgac aacctgcaga aggacttcaa gtccgccaag gacaccatca 1381 agaaacagat tagcgagtac atcaaggact ccgaaaagtt taaaaatctg ttcaaccaga 1441 atctgatcga tgctaagaaa ggccaggagt ccgacctgat cctgtggctg aaacagtcta 1501 aggacaatgg gattgaactg ttcaaggcta actccgatat cactgatatt gacgaggcac 1561 tggaaatcat caagagcttc aagggatgga ccacatactt taaaggcttc cacgagaacc 1621 gcaagaacgt gtactccagc aacgacattc ctacctccat catctaccga atcgtcgatg 1681 acaatctgcc aaagttcctg gagaacaagg ccaaatatga atctctgaag gacaaagctc 1741 ccgaggcaat taattacgaa cagatcaaga aagatctggc tgaggaactg acattcgata 1801 tcgactataa gactagcgag gtgaaccaga gggtcttttc cctggacgag gtgtttgaaa 1861 tcgccaattt caacaattac ctgaaccagt ccggcattac taaattcaat accatcattg 1921 gcgggaagtt tgtgaacggg gagaatacca agcgcaaggg aattaacgaa tacatcaatc 1981 tgtatagcca gcagatcaac gacaaaactc tgaagaaata caagatgtct gtgctgttca 2041 aacagatcct gagtgatacc gagtccaagt cttttgtcat tgataaactg gaagatgact 2101 cagacgtggt cactaccatg cagagctttt atgagcagat cgccgctttc aagacagtgg 2161 aggaaaaatc tattaaggaa actctgagtc tgctgttcga tgacctgaaa gcccagaagc 2221 tggacctgag taagatctac ttcaaaaacg ataagagtct gacagacctg tcacagcagg 2281 tgtttgatga ctattccgtg attgggaccg ccgtcctgga gtacattaca cagcagatcg 2341 ctccaaagaa cctggataat ccctctaaga aagagcagga actgatcgct aagaaaaccg 2401 agaaggcaaa atatctgagt ctggaaacaa ttaagctggc actggaggag ttcaacaagc 2461 acagggatat tgacaaacag tgccgctttg aggaaatcct ggccaacttc gcagccatcc 2521 ccatgatttt tgatgagatc gcccagaaca aagacaatct ggctcagatc agtattaagt 2581 accagaacca gggcaagaaa gacctgctgc aggcttcagc agaagatgac gtgaaagcca 2641 tcaaggatct gctggaccag accaacaatc tgctgcacaa gctgaaaatc ttccatatta 2701 gtcagtcaga ggataaggct aatatcctgg ataaagacga acacttctac ctggtgttcg 2761 aggaatgtta cttcgagctg gcaaacattg tccccctgta taacaagatt aggaactaca 2821 tcacacagaa gccttactct gacgagaagt ttaaactgaa cttcgaaaat agtaccctgg 2881 ccaacgggtg ggataagaac aaggagcctg acaacacagc tatcctgttc atcaaggatg 2941 acaagtacta tctgggagtg atgaataaga aaaacaataa gatcttcgat gacaaagcca 3001 ttaaggagaa caaaggggaa ggatacaaga aaatcgtgta taagctgctg cccggcgcaa 3061 ataagatgct gcctaaggtg ttcttcagcg ccaagagtat caaattctac aacccatccg 3121 aggacatcct gcggattaga aatcactcaa cacatactaa gaacgggagc ccccagaagg 3181 gatatgagaa atttgagttc aacatcgagg attgcaggaa gtttattgac ttctacaagc 3241 agagcatctc caaacaccct gaatggaagg attttggctt ccggttttcc gacacacaga 3301 gatataactc tatcgacgag ttctaccgcg aggtggaaaa tcaggggtat aagctgactt 3361 ttgagaacat ttctgaaagt tacatcgaca gcgtggtcaa tcagggaaag ctgtacctgt 3421 tccagatcta taacaaagat ttttcagcat acagcaaggg cagaccaaac ctgcatacac 3481 tgtactggaa ggccctgttc gatgagagga atctgcagga cgtggtctat aaactgaacg 3541 gagaggccga actgttttac cggaagcagt ctattcctaa gaaaatcact cacccagcta 3601 aggaggccat cgctaacaag aacaaggaca atcctaagaa agagagcgtg ttcgaatacg 3661 atctgattaa ggacaagcgg ttcaccgaag ataagttctt tttccattgt ccaatcacca 3721 ttaacttcaa gtcaagcggc gctaacaagt tcaacgacga gatcaatctg ctgctgaagg 3781 aaaaagcaaa cgatgtgcac atcctgagca ttgaccgagg agagcggcat ctggcctact 3841 ataccctggt ggatggcaaa gggaatatca ttaagcagga tacattcaac atcattggca 3901 atgaccggat gaaaaccaac taccacgata aactggctgc aatcgagaag gatagagact 3961 cagctaggaa ggactggaag aaaatcaaca acattaagga gatgaaggaa ggctatctga 4021 gccaggtggt ccatgagatt gcaaagctgg tcatcgaata caatgccatt gtggtgttcg 4081 aggatctgaa cttcggcttt aagagggggc gctttaaggt ggaaaaacag gtctatcaga 4141 agctggagaa aatgctgatc gaaaagctga attacctggt gtttaaagat aacgagttcg 4201 acaagaccgg aggcgtcctg agagcctacc agctgacagc tccctttgaa actttcaaga 4261 aaatgggaaa acagacaggc atcatctact atgtgccagc cggattcact tccaagatct 4321 gccccgtgac cggctttgtc aaccagctgt accctaaata tgagtcagtg agcaagtccc 4381 aggaattttt cagcaagttc gataagatct gttataatct ggacaagggg tacttcgagt 4441 tttccttcga ttacaagaac ttcggcgaca aggccgctaa ggggaaatgg accattgcct 4501 ccttcggatc tcgcctgatc aactttcgaa attccgataa aaaccacaat tgggacacta 4561 gggaggtgta cccaaccaag gagctggaaa agctgctgaa agactactct atcgagtatg 4621 gacatggcga atgcatcaag gcagccatct gtggcgagag tgataagaaa tttttcgcca 4681 agctgacctc agtgctgaat acaatcctgc agatgcggaa ctcaaagacc gggacagaac 4741 tggactatct gattagcccc gtggctgatg tcaacggaaa cttcttcgac agcagacagg 4801 cacccaaaaa tatgcctcag gatgcagacg ccaacggggc ctaccacatc gggctgaagg 4861 gactgatgct gctgggccgg atcaagaaca atcaggaggg gaagaagctg aacctggtca 4921 ttaagaacga ggaatacttc gagtttgtcc agaatagaaa taacaaaagg ccggcggcca 4981 cgaaaaaggc cggccaggca aaaaagaaaa agggatccta cccatacgat gttccagatt 5041 acgcttatcc ctacgacgtg cctgattatg catacccata tgatgtcccc gactatgccc 5101 caaagaagaa gcggaaggtc gatggttctg aaaacctgta cttccaatcc atgaaaatcg 5161 aagaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 5221 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 5281 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 5341 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 5401 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 5461 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 5521 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 5581 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 5641 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 5701 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 5761 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 5821 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 5881 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 5941 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 6001 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 6061 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ctatggaaaa 6121 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 6181 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 6241 cgcgcagact ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg 6301 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc 6361 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa 6421 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 6481 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 6541 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 6601 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 6661 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 6721 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 6781 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 6841 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa 6901 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 6961 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc 7021 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 7081 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 7141 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 7201 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 7261 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 7321 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 7381 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 7441 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 7501 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa 7561 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 7621 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 7681 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 7741 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 7801 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 7861 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 7921 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 7981 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 8041 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 8101 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 8161 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 8221 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 8281 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 8341 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 8401 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 8461 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 8521 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 8581 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 8641 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 8701 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 8761 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga 8821 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag 8881 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac 8941 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 9001 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 9061 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg 9121 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc 9181 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt 9241 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg 9301 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa 9361 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag 9421 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat 9481 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt 9541 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag 9601 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa 9661 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc 9721 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg 9781 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc 9841 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct 9901 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc 9961 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc 10021 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 10081 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 10141 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt 10201 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg 10261 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc 10321 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat 10381 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt 10441 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc 10501 tatcggctga atttgattgc gagtgagata tttatgccag ccagccaga //