LOCUS       Exported                4224 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBTBXh-4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4224)
  AUTHORS   Prior JE, Lynch MD, Gill RT
  TITLE     Broad-host-range vectors for protein expression across gram negative
            hosts.
  JOURNAL   Biotechnol Bioeng. 2010 Jun 1. 106(2):326-32.
  PUBMED    20148414
REFERENCE   2  (bases 1 to 4224)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4224
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             4..21
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      75..111
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     rep_origin      117..886
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             887..1549
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     CDS             1646..2836
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1998..2017)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      2856..2887
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2915..3793)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3820..4104
                     /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     4027..4046
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 aaccatcatc atcatcatca ttagaagctt gatatcattc aggacgagcc tcagactcca
       61 gcgtaactgg actgaaaaca aactaaagcg cccttgtggc gctttagttt tgttccgcgg
      121 ccaccggctg gctcgcttcg ctcggcccgt ggacaaccct gctggacaag ctgatggaca
      181 ggctgcgcct gcccacgagc ttgaccacag ggattgccca ccggctaccc agccttcgac
      241 cacataccca ccggctccaa ctgcgcggcc tgcggccttg ccccatcaat ttttttaatt
      301 ttctctgggg aaaagcctcc ggcctgcggc ctgcgcgctt cgcttgccgg ttggacacca
      361 agtggaaggc gggtcaaggc tcgcgcagcg accgcgcagc ggcttggcct tgacgcgcct
      421 ggaacgaccc aagcctatgc gagtgggggc agtcgaaggc gaagcccgcc cgcctgcccc
      481 ccgagcctca cggcggcgag tgcgggggtt ccaagggggc agcgccacct tgggcaaggc
      541 cgaaggccgc gcagtcgatc aacaagcccc ggaggggcca ctttttgccg gagggggagc
      601 cgcgccgaag gcgtggggga accccgcagg ggtgcccttc tttgggcacc aaagaactag
      661 atatagggcg aaatgcgaaa gacttaaaaa tcaacaactt aaaaaagggg ggtacgcaac
      721 agctcattgc ggcacccccc gcaatagctc attgcgtagg ttaaagaaaa tctgtaattg
      781 actgccactt ttacgcaacg cataattgtt gtcgcgctgc cgaaaagttg cagctgattg
      841 cgcatggtgc cgcaaccgtg cggcacccta ccgcatggag ataagcatgg ccacgcagtc
      901 cagagaaatc ggcattcaag ccaagaacaa gcccggtcac tgggtgcaaa cggaacgcaa
      961 agcgcatgag gcgtgggccg ggcttattgc gaggaaaccc acggcggcaa tgctgctgca
     1021 tcacctcgtg gcgcagatgg gccaccagaa cgccgtggtg gtcagccaga agacactttc
     1081 caagctcatc ggacgttctt tgcggacggt ccaatacgca gtcaaggact tggtggccga
     1141 gcgctggatc tccgtcgtga agctcaacgg ccccggcacc gtgtcggcct acgtggtcaa
     1201 tgaccgcgtg gcgtggggcc agccccgcga ccagttgcgc ctgtcggtgt tcagtgccgc
     1261 cgtggtggtt gatcacgacg accaggacga atcgctgttg gggcatggcg acctgcgccg
     1321 catcccgacc ctgtatccgg gcgagcagca actaccgacc ggccccggcg aggagccgcc
     1381 cagccagccc ggcattccgg gcatggaacc agacctgcca gccttgaccg aaacggagga
     1441 atgggaacgg cgcgggcagc agcgcctgcc gatgcccgat gagccgtgtt ttctggacga
     1501 tggcgagccg ttggagccgc cgacacgggt cacgctgccg cgccggtagt acgtaagagg
     1561 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     1621 attttcagga gctaaggaag ctaaaatgaa atctaacaat gcgctcatcg tcatcctcgg
     1681 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
     1741 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
     1801 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
     1861 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
     1921 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     1981 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     2041 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     2101 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
     2161 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat
     2221 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     2281 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     2341 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     2401 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     2461 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
     2521 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     2581 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     2641 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc
     2701 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     2761 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     2821 ggccacctcg acctgaatgg aagggatccc cctcaagtca aaagcctccg gtcggaggct
     2881 tttgactttc tgctatggag gtcaggtatg atttttatga caacttgacg gctacatcat
     2941 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
     3001 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
     3061 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
     3121 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
     3181 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
     3241 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
     3301 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
     3361 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
     3421 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
     3481 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
     3541 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
     3601 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
     3661 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
     3721 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     3781 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
     3841 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
     3901 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     3961 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     4021 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     4081 cgcaactctc tactgtttct ccatacccgt ttttttgggc tagcaccgcc tatctcgtgt
     4141 gagataggcg gagatacgaa ctttaagaag gagatatacc catggtaaaa tggtcagtat
     4201 tgagcgatat ctagagaatt cgtt
//
