LOCUS Exported 6176 bp ds-DNA circular SYN 13-OCT-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6176) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 1, 2017 from SnapGene 4.0.5 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6176 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 114..133 /label=Verification forward (VF2) primer binding site /note="/ApEinfo_fwdcolor=#85dae9 /ApEinfo_revcolor=#85dae9" terminator 184..227 /label=bacterial terminator /note="/ApEinfo_fwdcolor=#c6c9d1 /ApEinfo_revcolor=#c6c9d1" misc_feature 230..249 /label=BioBrick prefix CDS /label=BioBrick prefix /note="/Biobrick prefix for coding sequences /ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc" CDS 251..4357 /codon_start=1 /label=dCas9VRER /note="/ApEinfo_fwdcolor=#f58a5e /ApEinfo_revcolor=#f58a5e" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVL DATLIHQSITGLYETRIDLSQLGGD" misc_feature 278..280 /label=D10A /label=dCas9 /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd" misc_feature 1594 /label=T->A /note="/ApEinfo_fwdcolor=#f58a5e /ApEinfo_revcolor=#f58a5e" misc_feature 2768..2770 /label=H840A /label=dCas9 /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd" misc_feature 3629..4377 /label=gBlock_dCas9 misc_feature 3653..3655 /label=D1335V /note="/ApEinfo_fwdcolor=#85dae9 /ApEinfo_revcolor=#85dae9" misc_feature 3902..3904 /label=G1218R /note="/ApEinfo_fwdcolor=#c6c9d1 /ApEinfo_revcolor=#c6c9d1" misc_feature 4253..4255 /label=R1335E /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd" misc_feature 4259..4261 /label=T1337R /note="/ApEinfo_fwdcolor=#c7b0e3 /ApEinfo_revcolor=#c7b0e3" misc_feature 4358..4378 /label=BioBrick suffix /note="universal suffix for all parts" stem_loop 4379..4450 /label=E. coli his operon terminator /note="/ApEinfo_fwdcolor=#b7e6d7 /ApEinfo_revcolor=#b7e6d7" misc_feature complement(4514..4533) /label=Verification reverse (VR) primer binding site /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc" misc_feature complement(4618..5232) /label=rep (pMB1) /note="/ApEinfo_fwdcolor=#f8d3a9 /ApEinfo_revcolor=#f8d3a9" terminator 5402..5496 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(5517..6176) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" ORIGIN 1 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct 61 tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaactcga gtgccacctg 121 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 181 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgcggc 241 cgcttctaga atggataaga aatactcaat aggcttagct atcggcacaa atagcgtcgg 301 atgggcggtg atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa 361 tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga 421 gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa 481 tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt 541 ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc 601 tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca 661 tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc 721 cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga 781 taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga 841 agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag 901 taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt 961 atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga 1021 tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa 1081 tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc 1141 agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct 1201 atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc 1261 tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa 1321 cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata aatttatcaa 1381 accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa atcgtgaaga 1441 tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa ttcacttggg 1501 tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa aagacaatcg 1561 tgagaagatt gaaaaaatct tgacttttcg aataccttat tatgttggtc cattggcgcg 1621 tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta ccccatggaa 1681 ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac gcatgacaaa 1741 ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc tttatgagta 1801 ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc 1861 agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca aaacaaatcg 1921 aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat gttttgatag 1981 tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct accatgattt 2041 gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag atatcttaga 2101 ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg aaagacttaa 2161 aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc gccgttatac 2221 tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc aatctggcaa 2281 aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta tgcagctgat 2341 ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg 2401 cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta aaaaaggtat 2461 tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc ataagccaga 2521 aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc agaaaaattc 2581 gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa 2641 agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct attatctcca 2701 aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa gtgattatga 2761 tgtcgatgcc attgttccac aaagtttcct taaagacgat tcaatagaca ataaggtctt 2821 aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa 2881 aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc aacgtaagtt 2941 tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag ctggttttat 3001 caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa ttttggatag 3061 tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta aagtgattac 3121 cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata aagtacgtga 3181 gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg gaactgcttt 3241 gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata aagtttatga 3301 tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg caaaatattt 3361 cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa atggagagat 3421 tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct gggataaagg 3481 gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa 3541 aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga 3601 caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt ttgttagtcc 3661 aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt 3721 aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa 3781 tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa 3841 actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc 3901 ccgagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata 3961 tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt 4021 gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc 4081 taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca 4141 tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa 4201 tcttggagct cccgctgctt ttaaatattt tgatacaaca attgatcgta aagaatatcg 4261 ttctacaaaa gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga 4321 aacacgcatt gatttgagtc agctaggagg tgactgatac tagtagcggc cgctgcagtc 4381 cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac 4441 ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 4501 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 4561 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 4621 tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 4681 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 4741 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 4801 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 4861 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 4921 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 4981 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 5041 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 5101 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 5161 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5221 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5281 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5341 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc tcgaggcttg 5401 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga 5461 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac 5521 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg 5581 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct 5641 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg 5701 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca 5761 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat 5821 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca 5881 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg 5941 tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata 6001 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc 6061 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct 6121 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccat //