LOCUS Exported 5742 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBAD His6 TEV LIC transfer vector (8BT) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5742) TITLE 8-series E. coli arabinose expression vectors JOURNAL Unpublished REFERENCE 2 (bases 1 to 5742) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5742 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 858..1446 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1347..1366 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1600..1617 /label=L4440 /note="L4440 vector, forward primer" misc_feature 1632..1772 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(1698..1717) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1858..1880 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(1874..2065) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 3523..3542 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(3701..4579) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 4606..4890 /gene="araBAD" /label=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)" primer_bind 4813..4832 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 4953..4975 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4995..5012 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 5013..5033 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" primer_bind complement(5123..5140) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 5293..5339 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(5379..5397) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5465..5569 /gene="bla" /label=AmpR promoter CDS 5570..688 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" ORIGIN 1 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 61 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 121 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 181 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 241 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 301 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 361 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 421 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 481 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 541 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 601 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 661 aggtgcctca ctgattaagc attggtaacc cgggaccaag tttactcata tatactttag 721 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 781 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 841 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 901 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 961 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 1021 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 1081 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 1141 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 1201 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 1261 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 1321 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 1381 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 1441 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 1501 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 1561 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 1621 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 1681 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc 1741 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg 1801 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 1861 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt 1921 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca 1981 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa 2041 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg 2101 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac 2161 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc 2221 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac 2281 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact 2341 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg 2401 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct 2461 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca 2521 tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga 2581 tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca 2641 agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc 2701 cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag 2761 gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata 2821 aatcgccgtg acgatcagcg gtccagtgat cgaagttagg ctggtaagag ccgcgagcga 2881 tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc 2941 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg 3001 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc 3061 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg 3121 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc 3181 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa 3241 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg 3301 gttgaaggct ctcaagggca tcggtcgacg ctctccctta tgcgactcct gcattaggaa 3361 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa 3421 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca 3481 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata 3541 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag 3601 gatcgcgacc gccataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat 3661 gctactccct cgagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta 3721 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt 3781 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg 3841 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc 3901 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg 3961 acaagcaaac atgctgtgcg acgctggcta tatcaaaatt gctgtctgcc aggtgatcgc 4021 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa 4081 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat 4141 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct 4201 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa gattgacggc cagttaagcc 4261 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct 4321 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc 4381 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga 4441 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct 4501 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat 4561 cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc 4621 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac 4681 cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa 4741 cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg 4801 tcacactttg ctatgccata gcatttttat ccataagatt agcggatctt acctgacgct 4861 ttttatcgca actctctact gtttctccat acccgttttt ttgggctatt aattaagcgg 4921 ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa ggagatatac 4981 atatgaaatc ttctcaccat caccatcacc atgaaaacct gtacttccaa tccaatattg 5041 gaagtggata acggatccgc gatcgcggcg cgccacctgg tggccggcct ggctgttttg 5101 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 5161 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 5221 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 5281 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 5341 tgttgtttgt cggtgaacta attcttgaag acgaaagggc ctcgtgatac gcctattttt 5401 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa 5461 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 5521 gagacaataa ccctgataaa tgcttcaata acattgaaaa aggaagagta tgagtattca 5581 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 5641 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 5701 catcgaactg gatctcaaca gcggtaagat ccttgagagt tt //