LOCUS       sequence-12242-e        3845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3845)
  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 3845)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            Bioeng. 2010 Jun 1. 106(2):326-32."
COMMENT     Sequence Label: pBTBXh-2
FEATURES             Location/Qualifiers
     source          1..3845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..21
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      complement(74..110)
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     rep_origin      116..885
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             886..1548
                     /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             1645..2460
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1718..1737)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2477..2508)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2536..3414)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3441..3725
                     /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     3648..3667
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 gaccatcatc atcatcatca ttagagcttg atatcattca ggacgagcct cagactccag
       61 cgtaactgga ctgaaaacaa actaaagcgc ccttgtggcg ctttagtttt gttccgcggc
      121 caccggctgg ctcgcttcgc tcggcccgtg gacaaccctg ctggacaagc tgatggacag
      181 gctgcgcctg cccacgagct tgaccacagg gattgcccac cggctaccca gccttcgacc
      241 acatacccac cggctccaac tgcgcggcct gcggccttgc cccatcaatt tttttaattt
      301 tctctgggga aaagcctccg gcctgcggcc tgcgcgcttc gcttgccggt tggacaccaa
      361 gtggaaggcg ggtcaaggct cgcgcagcga ccgcgcagcg gcttggcctt gacgcgcctg
      421 gaacgaccca agcctatgcg agtgggggca gtcgaaggcg aagcccgccc gcctgccccc
      481 cgagcctcac ggcggcgagt gcgggggttc caagggggca gcgccacctt gggcaaggcc
      541 gaaggccgcg cagtcgatca acaagccccg gaggggccac tttttgccgg agggggagcc
      601 gcgccgaagg cgtgggggaa ccccgcaggg gtgcccttct ttgggcacca aagaactaga
      661 tatagggcga aatgcgaaag acttaaaaat caacaactta aaaaaggggg gtacgcaaca
      721 gctcattgcg gcaccccccg caatagctca ttgcgtaggt taaagaaaat ctgtaattga
      781 ctgccacttt tacgcaacgc ataattgttg tcgcgctgcc gaaaagttgc agctgattgc
      841 gcatggtgcc gcaaccgtgc ggcaccctac cgcatggaga taagcatggc cacgcagtcc
      901 agagaaatcg gcattcaagc caagaacaag cccggtcact gggtgcaaac ggaacgcaaa
      961 gcgcatgagg cgtgggccgg gcttattgcg aggaaaccca cggcggcaat gctgctgcat
     1021 cacctcgtgg cgcagatggg ccaccagaac gccgtggtgg tcagccagaa gacactttcc
     1081 aagctcatcg gacgttcttt gcggacggtc caatacgcag tcaaggactt ggtggccgag
     1141 cgctggatct ccgtcgtgaa gctcaacggc cccggcaccg tgtcggccta cgtggtcaat
     1201 gaccgcgtgg cgtggggcca gccccgcgac cagttgcgcc tgtcggtgtt cagtgccgcc
     1261 gtggtggttg atcacgacga ccaggacgaa tcgctgttgg ggcatggcga cctgcgccgc
     1321 atcccgaccc tgtatccggg cgagcagcaa ctaccgaccg gccccggcga ggagccgccc
     1381 agccagcccg gcattccggg catggaacca gacctgccag ccttgaccga aacggaggaa
     1441 tgggaacggc gcgggcagca gcgcctgccg atgcccgatg agccgtgttt tctggacgat
     1501 ggcgagccgt tggagccgcc gacacgggtc acgctgccgc gccggtagta cgtaagaggt
     1561 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     1621 ttttcaggag ctaaggaagc taaaatgagc catattcaac gggaaacgtc ttgctcgagg
     1681 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1741 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     1801 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcaggcta
     1861 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1921 gatgcatggt tactcaccac tgcgatccca gggaaaacag cattccaggt attagaagaa
     1981 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     2041 tcgattcctg tttgtaattg tccttttaac ggcgatcgcg tatttcgtct cgctcaggcg
     2101 caatcacgaa tgaataacgg tttggttggt gcgagtgatt ttgatgacga gcgtaatggc
     2161 tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca
     2221 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     2281 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2341 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2401 attgataatc ctgatatgaa taaattgcag tttcacttga tgctcgatga gtttttctga
     2461 gggcggatcc ccctcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     2521 ggtcaggtat gatttttatg acaacttgac ggctacatca ttcacttttt cttcacaacc
     2581 ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag
     2641 ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa
     2701 aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa
     2761 ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct
     2821 ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac
     2881 ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa
     2941 ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt
     3001 aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa
     3061 ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg
     3121 aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat
     3181 ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt
     3241 tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg
     3301 tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag
     3361 atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt
     3421 catactcccg ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca
     3481 ctgcgtcttt tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat
     3541 tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca
     3601 cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt
     3661 tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc
     3721 tccatacccg tttttttggg ctagcaccgc ctatctcgtg tgagataggc ggagatacga
     3781 actttaagaa ggagatatac ccatggtaaa atggtcagta ttgagcgata tctagagaat
     3841 tcgtc
//
