LOCUS Exported 5975 bp ds-DNA circular SYN 16-MAR-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Untitled 5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5975) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, Mar 16, 2017 from SnapGene Viewer 3.2.1 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5975 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..285 /gene="araBAD" /note="araBAD promoter" /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" CDS 331..348 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 352..384 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /note="T7 tag (gene 10 leader)" /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 388..411 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /note="Xpress(TM) tag" /translation="DLYDDDDK" CDS 427..1152 /codon_start=1 /note="PhoCl" /translation="VIPDYFKQSFPEGYSWERSMTYEDGGICIATNDITMEGDSFINKI HFKGTNFPPNGPVMQKRTVGWEASTEKMYERDGVLKGDVKMKLLLKGGGHYRCDYRTTY KVKQKPVKLPDYHFVDHRIEILSHDKDYNKVKLYEHAVARNSTDSMDELYKGGSGGMVS KGEETITSVIKPDMKNKLRMEGNVNGHAFVIEGEGSGKPFEGIQTIDLEVKEGAPLPFA YDILTTAFHYGNRVFTKYPR" CDS 1168..2268 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /note="MBP" /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" CDS 2317..2328 /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" terminator 2546..2632 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2724..2751 /note="rrnB T2 terminator" /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2770..2861 /gene="bla" /note="AmpR promoter" CDS 2862..3722 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3893..4481 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 4667..4807 /note="bom" /note="basis of mobility region from pBR322" CDS complement(5071..5949) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /note="araC" /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct 61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca 121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg 181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg 241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc 301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc 361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc 421 tcgagtgtga tccctgacta cttcaagcag agcttccccg agggctacag ctgggagcgc 481 agcatgacct acgaggacgg cggcatctgc atcgccacca acgacatcac aatggagggg 541 gacagcttca tcaacaagat ccacttcaag ggcacgaact tcccccccaa cggccccgtg 601 atgcagaaga ggaccgtggg ctgggaggcc agcaccgaga agatgtacga gcgcgacggc 661 gtgctgaagg gcgacgtgaa gatgaagctg ctgctgaagg gcggcggcca ctatcgctgc 721 gactaccgca ccacctacaa ggtcaagcag aagcccgtaa agctgcccga ctaccacttc 781 gtggaccacc gcatcgagat cctgagccac gacaaggact acaacaaggt gaagctgtac 841 gagcacgccg tggcccgcaa ctccaccgac agcatggacg agctgtacaa gggtggcagc 901 ggtggcatgg tgagcaaggg cgaggagacc attacaagcg tgatcaagcc tgacatgaag 961 aacaagctgc gcatggaggg caacgtgaac ggccacgcct tcgtgatcga gggcgagggc 1021 agcggcaagc ccttcgaggg catccagacg attgatttgg aggtgaagga gggcgccccg 1081 ctgcccttcg cctacgacat cctgaccacc gccttccact acggcaaccg cgtgttcacc 1141 aagtacccac ggggaggtgg aggtaccatg aaaatcgaag aaggtaaact ggtaatctgg 1201 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 1261 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 1321 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 1381 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 1441 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 1501 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 1561 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 1621 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 1681 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 1741 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 1801 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 1861 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 1921 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 1981 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2041 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2101 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg 2161 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2221 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2281 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcataaaag 2341 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc 2401 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac 2461 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat 2521 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc 2581 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg 2641 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata 2701 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct 2761 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 2821 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 2881 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 2941 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 3001 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 3061 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag 3121 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 3181 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 3241 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 3301 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 3361 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 3421 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 3481 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 3541 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 3601 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 3661 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 3721 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 3781 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 3841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 3901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 3961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 4021 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact 4081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 4141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 4201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 4261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 4321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 4381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 4441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 4501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 4561 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc 4621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt 4681 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct 4741 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat 4801 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 4861 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 4921 accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca 4981 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt 5041 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac 5101 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac 5161 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat 5221 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa 5281 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac 5341 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg 5401 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat 5461 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc 5521 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc 5581 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca 5641 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg 5701 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa 5761 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc 5821 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt 5881 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc 5941 ttcagccata cttttcatac tcccgccatt cagag //