LOCUS Exported 5742 bp ds-DNA circular SYN 26-JAN-2023 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synthetic DNA construct REFERENCE 1 (bases 1 to 5742) TITLE 8-series E. coli arabinose expression vectors JOURNAL Unpublished REFERENCE 2 (bases 1 to 5742) AUTHORS .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 Jan 26, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5742 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 21..67 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 220..237 /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" CDS complement(327..347) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS complement(348..365) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" RBS 385..407 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" promoter complement(470..754) /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 complement(528..547) /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 781..1659 /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" primer_bind complement(1818..1837) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS 3295..3486 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind complement(3480..3502) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3588..3728 /label=bom /note="basis of mobility region from pBR322" primer_bind 3643..3662 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3743..3760) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3914..4502) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3994..4013) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4672..5532) /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" primer_bind 5295..5314 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5533..5637) /gene="bla" /label=AmpR promoter primer_bind 5705..5723 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 agttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt 61 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg 121 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg 181 ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg 241 aaaatcttct ctcatccgcc aaaacagcca ggccggccac caggtggcgc gccgcgatcg 301 cggatccgtt atccacttcc aatattggat tggaagtaca ggttttcatg gtgatggtga 361 tggtgagaag atttcatatg tatatctcct tcttaaagtt aaacaaaatt atttctagaa 421 ctcgagtcct gcagggcggc cgcttaatta atagcccaaa aaaacgggta tggagaaaca 481 gtagagagtt gcgataaaaa gcgtcaggta agatccgcta atcttatgga taaaaatgct 541 atggcatagc aaagtgtgac gccgtgcaaa taatcaatgt ggacttttct gccgtgatta 601 tagacacttt tgttacgcgt ttttgtcatg gctttggtcc cgctttgtta cagaatgctt 661 ttaataagcg gggttaccgg tttggttagc gagaagagcc agtaaaagac gcagtgacgg 721 caatgtctga tgcaatatgg acaattggtt tcttctctga atggcgggag tatgaaaagt 781 atggctgaag cgcaaaatga tcccctgctg ccgggatact cgtttaatgc ccatctggtg 841 gcgggtttaa cgccgattga ggccaacggt tatctcgatt tttttatcga ccgaccgctg 901 ggaatgaaag gttatattct caatctcacc attcgcggtc agggggtggt gaaaaatcag 961 ggacgagaat ttgtttgccg accgggtgat attttgctgt tcccgccagg agagattcat 1021 cactacggtc gtcatccgga ggctcgcgaa tggtatcacc agtgggttta ctttcgtccg 1081 cgcgcctact ggcatgaatg gcttaactgg ccgtcaatct ttgccaatac ggggttcttt 1141 cgcccggatg aagcgcacca gccgcatttc agcgacctgt ttgggcaaat cattaacgcc 1201 gggcaagggg aagggcgcta ttcggagctg ctggcgataa atctgcttga gcaattgttg 1261 ctgcggcgca tggaagcgat taacgagtcg ctccatccac cgatggataa tcgggtacgc 1321 gaggcttgtc agtacatcag cgatcacctg gcagacagca attttgatat agccagcgtc 1381 gcacagcatg tttgcttgtc gccgtcgcgt ctgtcacgtc ttttccgcca gcagttaggg 1441 attagcgtct taagctggcg cgaggaccaa cgtatcagcc aggcgaagct gcttttgagc 1501 accacccgga tgcctatcgc caccgtcggt cgcaatgttg gttttgacga tcaactctat 1561 ttctcgcggg tatttaaaaa atgcaccggg gccagcccga gcgagttccg tgccggttgt 1621 gaagaaaaag tgaatgatgt agccgtcaag ttgtcataat tggtaacgaa tcagacaatt 1681 gacggctcga gggagtagca tagggtttgc agaatccctg cttcgtccat ttgacgggca 1741 cattatggcg gtcgcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac 1801 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca 1861 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg 1921 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct 1981 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat 2041 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt 2101 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct 2161 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct 2221 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa 2281 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt 2341 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc 2401 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca 2461 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcattggacc 2521 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat 2581 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg 2641 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga 2701 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac 2761 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg 2821 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg 2881 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc 2941 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg 3001 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca 3061 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga 3121 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa 3181 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt 3241 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc 3301 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt 3361 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac 3421 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac 3481 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 3541 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc 3601 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg 3661 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 3721 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 3781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 3841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 3901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 3961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4321 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 4381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 4441 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 4501 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 4561 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 4621 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtcccgg gttaccaatg 4681 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 4741 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 4801 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 4861 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 4921 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 4981 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 5041 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 5101 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 5161 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 5221 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 5281 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 5341 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 5401 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 5461 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 5521 ttgaatactc atactcttcc tttttcaatg ttattgaagc atttatcagg gttattgtct 5581 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 5641 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 5701 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa tt //