LOCUS Exported 7572 bp ds-DNA circular SYN 13-JUN-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDG1662-sgRNA_071515 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7572) AUTHORS Jason X. M. Peters TITLE Direct Submission JOURNAL Exported Monday, Jun 13, 2016 from SnapGene 3.1.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7572 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 115..635 /codon_start=1 /transl_table=11 /gene="amyE" /product="alpha-amylase" /note="Bsu amyE'" /note="amy-front" /db_xref="GI:1185566" /protein_id="AAB40299.1" /translation="MFAKRFKTSLLPLFAGFLLLFHLVLAGPAAASAETANKSNELTAP SIKSGTILHAWNWSFNTLKHNMKDIHDAGYTAIQTSPINQVKEGNQGDKSMSNWYWLYQ PTSYQIGNRYLGTEQEFKEMCAAAEEYGIKVIVDAVINHTTSDYAAISNEVKSIPNWTH GNTQIKNWSD" promoter 660..724 /note="Bsu veg promoter" misc_feature 725..744 /note="RR1" /note="mrfp sgRNA sequence, also called ""NT1""" misc_feature 745..827 /note="sgRNA" /note="single guide RNA" CDS 841..870 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /note="Myc" /translation="EQKLISEEDL" CDS 886..903 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" terminator 1121..1184 /note="rrnB T1 terminator" CDS 1556..2206 /codon_start=1 /transl_table=11 /gene="cat" /product="chloramphenicol acetyltransferase" /note="cat" /note="Gram positive inducible" /db_xref="GI:20804332" /experiment="experimental evidence, no additional details recorded" /protein_id="CAD10262.1" /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL" gene 1556..2206 /gene="cat" /note="cat" CDS 2577..3605 /codon_start=1 /transl_table=11 /gene="amyE" /product="alpha-amylase" /note="Bsu 'amyE" /note="amy-back" /db_xref="GI:1185568" /protein_id="AAB40301.1" /translation="STWMSDDDIRLGWAVIASRSGSTPLFFSRPEGGGNGVRFPGKSQI GDRGSALFEDQAITAVNRFHNVMAGQPEELSNPNGNNQIFMNQRGSHGVVLANAGSSSV SINTATKLPDGRYDNKAGAGSFQVNDGKLTGTINARSVAVLYPDDIAKAPHVFLENYKT GVTHSFNDQLTITLRADANTTKAVYQINNGPDDRRLRMEINSQSEKEIQFGKTYTIMLK GTNSDGVTRTEKYSFVKRDPASAKTIGYQNPNHWSQVNAYIYKHDGSRVIELTGSWPGK PMTKNADGIYTLTLPADTDTTNAKVIFNNGSAQVPGQNQPGFDYVLNGLYNDSGLSGSL PH" gene 4714..5496 /gene="spc" /note="spc" CDS 4714..5496 /codon_start=1 /transl_table=11 /gene="spc" /product="spectinomycin adenyltransferase" /note="spc" /note="spectinomycin resistance" /db_xref="GI:1185569" /protein_id="AAB40302.1" /translation="MSNLINGKIPNQAIQTLKIVKDLFGSSIVGVYLFGSAVNGGLRIN SDVDVLVVVNHSLPQLTRKKLTERLMTISGKIGNTDSVRPLEVTVINRSEVVPWQYPPK REFIYGEWLRGEFENGQIQEPSYDPDLAIVLAQARKNSISLFGPDSSSILVSVPLTDIR RAIKDSLPELIEGIKGDERNVILTLARMWQTVTTGEITSKDVAAEWAIPLLPKEHVTLL DIARKGYRGECDDKWEGLYSKVKALVKYMKNSIETSLN" rep_origin complement(5743..6331) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6502..7362) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" gene complement(6502..7362) /gene="bla" /note="bla" promoter complement(7363..7467) /gene="bla" /note="AmpR promoter" ORIGIN 1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga 61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt 121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat 181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt 241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg 301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg 361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg 421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa 481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat 541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca 601 catggaaaca cacaaattaa aaactggtct gatcggatcc tagaagctta tcgaattcct 661 tattaacgtt gatataattt aaattttatt tgacaaaaat gggctcgtgt tgtacaataa 721 atgtaacttt cagtttagcg gtctgtttta gagctagaaa tagcaagtta aaataaggct 781 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgaa gcttgggccc 841 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat 901 cattgagttt aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc 961 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca 1021 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg 1081 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga 1141 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaactggatc 1201 cttactcgag tctagactgc aggcttcctc gctcactgac tcgaattctc atgtttgaca 1261 gcttatcatc ggcaatagtt acccttatta tcaagataag aaagaaaagg atttttcgct 1321 acgctcaaat cctttaaaaa aacacaaaag accacatttt ttaatgtggt ctttattctt 1381 caactaaagc acccattagt tcaacaaacg aaaattggat aaagtgggat atttttaaaa 1441 tatatattta tgttacagta atattgactt ttaaaaaagg attgattcta atgaagaaag 1501 cagacaagta agcctcctaa attcacttta gataaaaatt taggaggcat atcaaatgaa 1561 ctttaataaa attgatttag acaattggaa gagaaaagag atatttaatc attatttgaa 1621 ccaacaaacg acttttagta taaccacaga aattgatatt agtgttttat accgaaacat 1681 aaaacaagaa ggatataaat tttaccctgc atttattttc ttagtgacaa gggtgataaa 1741 ctcaaataca gcttttagaa ctggttacaa tagcgacgga gagttaggtt attgggataa 1801 gttagagcca ctttatacaa tttttgatgg tgtatctaaa acattctctg gtatttggac 1861 tcctgtaaag aatgacttca aagagtttta tgatttatac ctttctgatg tagagaaata 1921 taatggttcg gggaaattgt ttcccaaaac acctatacct gaaaatgctt tttctctttc 1981 tattattcca tggacttcat ttactgggtt taacttaaat atcaataata atagtaatta 2041 ccttctaccc attattacag caggaaaatt cattaataaa ggtaattcaa tatatttacc 2101 gctatcttta caggtacatc attctgtttg tgatggttat catgcaggat tgtttatgaa 2161 ctctattcag gaattgtcag ataggcctaa tgactggctt ttataatatg agataatgcc 2221 gactgtactt tttacagtcg gttttctaat gtcactaacc tgccccgtta gttgaagaag 2281 gtttttatat tacagctcca gatcctctac gccggacgca tcgtggccgg catcaccggc 2341 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct 2401 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc 2461 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac 2521 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga 2581 catggatgag cgatgatgat atccgtttag gctgggcggt gatagcttct cgttcaggca 2641 gtacgcctct tttcttttcc agacctgagg gaggcggaaa tggtgtgagg ttcccgggga 2701 aaagccaaat aggcgatcgc gggagtgctt tatttgaaga tcaggctatc actgcggtca 2761 atagatttca caatgtgatg gctggacagc ctgaggaact ctcgaacccg aatggaaaca 2821 accagatatt tatgaatcag cgcggctcac atggcgttgt gctggcaaat gcaggttcat 2881 cctctgtctc tatcaatacg gcaacaaaat tgcctgatgg caggtatgac aataaagctg 2941 gagcgggttc atttcaagtg aacgatggta aactgacagg cacgatcaat gccaggtctg 3001 tagctgtgct ttatcctgat gatattgcaa aagcgcctca tgttttcctt gagaattaca 3061 aaacaggtgt aacacattct ttcaatgatc aactgacgat taccttgcgt gcagatgcga 3121 atacaacaaa agccgtttat caaatcaata atggaccaga cgacaggcgt ttaaggatgg 3181 agatcaattc acaatcggaa aaggagatcc aatttggcaa aacatacacc atcatgttaa 3241 aaggaacgaa cagtgatggt gtaacgagga ccgagaaata cagttttgtt aaaagagatc 3301 cagcgtcggc caaaaccatc ggctatcaaa atccgaatca ttggagccag gtaaatgctt 3361 atatctataa acatgatggg agccgagtaa ttgaattgac cggatcttgg cctggaaaac 3421 caatgactaa aaatgcagac ggaatttaca cgctgacgct gcctgcggac acggatacaa 3481 ccaacgcaaa agtgattttt aataatggca gcgcccaagt gcccggtcag aatcagcctg 3541 gctttgatta cgtgctaaat ggtttatata atgactcggg cttaagcggt tctcttcccc 3601 attgagggca aggctagacg ggacttaccg aaagaaacca tcaatgatgg tttctttttt 3661 gttcataaat cagacaaaac ttttctcttg caaaagtttg tgaagtgttg cacaatataa 3721 atgtgaaata cttcacaaac aaaaagacat caaagagaaa cataccctgc aaggatgctg 3781 atattgtctg catttgcgcc ggagcaaacc aaaaacctgg tgagacacgc cttgaattag 3841 tagaaaagaa cttgaagatt ttcaaaggca tcgttagtga agtcatggcg agcggatttg 3901 acggcatttt cttagtcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg 3961 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg 4021 accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg 4081 gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat 4141 ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga 4201 gccgggccac ctactgaagt ggatttcttt aagagctcct ttaacttcct caccagtagt 4261 tgtatcggta ccataagtag aagcagcaac ccaagtagct ttaccagcat ccggttcaac 4321 cagcatagta agaatcttac tggacatcgg cagttcttcg aacagtgcgc caactaccag 4381 ctctttctgc agttcattca gggcaccgga gaacctgcgt gcaatccatc ttgttcaatc 4441 atgcgaaacg atcctcatcc tgtctcttga tccatggatt acgcgttaac ccgggcccgc 4501 ggatgcatat gatcagatct taaggcctag gtctagagga tcgatctgta taataaagaa 4561 taattattaa tctgtagaca aattgtgaaa ggatgtactt aaacgctaac ggtcagcttt 4621 attgaacagt aatttaagta tatgtccaat ctagggtaag taaattgagt atcaatataa 4681 actttatatg aacataatca acgaggtgaa atcatgagca atttgattaa cggaaaaata 4741 ccaaatcaag cgattcaaac attaaaaatc gtaaaagatt tatttggaag ttcaatagtt 4801 ggagtatatc tatttggttc agcagtaaat ggtggtttac gcattaacag cgatgtagat 4861 gttctagtcg tcgtgaatca tagtttacct caattaactc gaaaaaaact aacagaaaga 4921 ctaatgacta tatcaggaaa gattggaaat acggattctg ttagaccact tgaagttacg 4981 gttataaata ggagtgaagt tgtcccttgg caatatcctc caaaaagaga atttatatac 5041 ggtgagtggc tcaggggtga atttgagaat ggacaaattc aggaaccaag ctatgatcct 5101 gatttggcta ttgttttagc acaagcaaga aagaatagta tttctctatt tggtcctgat 5161 tcttcaagta tacttgtctc cgtacctttg acagatattc gaagagcaat taaggattct 5221 ttgccagaac taattgaggg gataaaaggt gatgagcgta atgtaatttt aaccctagct 5281 cgaatgtggc aaacagtgac tactggtgaa attacctcga aagatgtcgc tgcagaatgg 5341 gctatacctc ttttacctaa agagcatgta actttactgg atatagctag aaaaggctat 5401 cggggagagt gtgatgataa gtgggaagga ctatattcaa aggtgaaagc actcgttaag 5461 tatatgaaaa attctataga aacttctctc aattaggcta attttattgc aataacaggt 5521 gcttactttt ctggagttct ttagcaaatt tttttattag ctgaacttag tattagtggc 5581 catactcctc caatccaaag ctatttagaa agattactat atcctcaaac aggcggtaac 5641 cggcctcttc atcgggaatg cgcgcgacct tcagcatcgc cggcatgtcc ccctggcgga 5701 cgggaagtat ccagctcgag gtcgggccgc gttgctggcg tttttccata ggctccgccc 5761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 5821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 5881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 5941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6181 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttga 6241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 6361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 6421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 6481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 6541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 6601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 6661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 6721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 6781 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc 6841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 6901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 6961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca 7141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 7381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 7441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 7501 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 7561 tcttcaagaa tt //