LOCUS       sequence-254851-        5659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host range vector with toxin-antitoxin stability system for 
            maintenance in the absence of antibiotic selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5659)
  AUTHORS   Burbank L, Wei W
  TITLE     Broad-Host-Range Plasmids for Constitutive and Inducible Gene 
            Expression in the Absence of Antibiotic Selection.
  JOURNAL   Microbiol Resour Announc. 2019 Sep 5;8(36). pii: 8/36/e00769-19. 
            doi: 10.1128/MRA.00769-19.
  PUBMED    31488530
REFERENCE   2  (bases 1 to 5659)
  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: "Microbiol 
            Resour Announc."; date: "2019-09-5"; volume: "8(36). pii: 
            8/36/e00769-19. doi"; pages: " 10.1128/MRA.00769-19"
COMMENT     Sequence Label: pBBR4pemIK
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      66..837
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             838..1500
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     2194..2216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2208..2225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2209..2225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2235..2254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2235..2253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2286..2302
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2286..2302
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3041..3057)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3042..3058)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3087..3107)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3087..3105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3126..3142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3126..3142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3139..3161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3150..3166
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3174..3180,3181..3198,3199..3204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3219..3240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3468..3539
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3540..3608,3609..4400)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3758..3777)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggcttcgccc tgcgcggtcg ctgcgctccc ttgccagccc gtggatatgt ggacgatggc
       61 cgcgagcggc caccggctgg ctcgcttcgc tcggcccgtg gacaaccctg ctggacaagc
      121 tgatggacag gctgcgcctg cccacgagct tgaccacagg gattgcccac cggctaccca
      181 gccttcgacc acatacccac cggctccaac tgcgcggcct gcggccttgc cccatcaatt
      241 tttttaattt tctctgggga aaagcctccg gcctgcggcc tgcgcgcttc gcttgccggt
      301 tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga ccgcgcagcg gcttggcctt
      361 gacgcgcctg gaacgaccca agcctatgcg agtgggggca gtcgaagggc gaagcccgcc
      421 cgcctgcccc ccgagcctca cggcggcgag tgcgggggtt ccaagggggc agcgccacct
      481 tgggcaaggc cgaaggccgc gcagtcgatc aacaagcccc ggaggggcca ctttttgccg
      541 gagggggagc cgcgccgaag gcgtggggga accccgcagg ggtgcccttc tttgggcacc
      601 aaagaactag atatagggcg aaatgcgaaa gacttaaaaa tcaacaactt aaaaaagggg
      661 ggtacgcaac agctcattgc ggcacccccc gcaatagctc attgcgtagg ttaaagaaaa
      721 tctgtaattg actgccactt ttacgcaacg cataattgtt gtcgcgctgc cgaaaagttg
      781 cagctgattg cgcatggtgc cgcaaccgtg cggcacccct accgcatgga gataagcatg
      841 gccacgcagt ccagagaaat cggcattcaa gccaagaaca agcccggtca ctgggtgcaa
      901 acggaacgca aagcgcatga ggcgtgggcc gggcttattg cgaggaaacc cacggcggca
      961 atgctgctgc atcacctcgt ggcgcagatg ggccaccaga acgccgtggt ggtcagccag
     1021 aagacacttt ccaagctcat cggacgttct ttgcggacgg tccaatacgc agtcaaggac
     1081 ttggtggccg agcgctggat ctccgtcgtg aagctcaacg gccccggcac cgtgtcggcc
     1141 tacgtggtca atgaccgcgt ggcgtggggc cagccccgcg accagttgcg cctgtcggtg
     1201 ttcagtgccg ccgtggtggt tgatcacgac gaccaggacg aatcgctgtt ggggcatggc
     1261 gacctgcgcc gcatcccgac cctgtatccg ggcgagcagc aactaccgac cggccccggc
     1321 gaggagccgc ccagccagcc cggcattccg ggcatggaac cagacctgcc agccttgacc
     1381 gaaacggagg aatgggaacg gcgcgggcag cagcgcctgc cgatgcccga tgagccgtgt
     1441 tttctggacg atggcgagcc gttggagccg ccgacacggg tcacgctgcc gcgccggtag
     1501 cacttgggtt gcgcagcaac ccgtaagtgc gctgttccag actatcggct gtagccgcct
     1561 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     1621 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccgtttt tatcaggctc
     1681 tgggaggcag aataaatgat catatcgtca attattacct ccacggggag agcctgagca
     1741 aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     1801 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     1861 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     1921 aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     1981 catcgcagta cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2041 tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg
     2101 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2161 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2221 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg
     2281 gccgctctag aactagtgga tcccccgggc tgcaggaatt cactagtgat tggttgcagt
     2341 ggcctgaact tgccccaaaa attactggaa gatggtgaca gcttttgcca agacatcggc
     2401 caagatgttt ggtggtaggc tttccacttt gcgggcattg cgagcgagta agtcgagcgc
     2461 acgtggttga tcacatcgaa cgacgcccgt tgtaatcgta cctagtaatt ccacagcgaa
     2521 cccgaggcgg gaggcgaact cgccgccgtt tgtgattgga aggatcaccg gacaagaggt
     2581 aagccgattg aatcgctcac tggagataac aaccaccgga cggtgaccgc gttgttcgtg
     2641 tccggcggtt ggctccagat cgaccatgta aacatcgccg cgtttcatat cagttcgcgt
     2701 cctactgctg gagcatcgac ccattcacgt tcctcgggcg gcaacgggta agccccggag
     2761 gcttccgctt cggccagcag ttgatccagc gtgtactgag gatgtaggac tggctcaatg
     2821 accagacgac cgtctgtgac ggctaagccg accgttgcgc cgacctcaag atgaagctga
     2881 tcaaggaaag cggggggaac ggccagcatg atcgatccac cgactttacg aaggtttgtg
     2941 gtgtgcatga tgggcttctt tcgttaagtt atattaaagt ataatataat tcctccattt
     3001 atacgcttat agcgaaaatc gaattcgata tcaagcttat cgataccgtc gacctcgagg
     3061 gggggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     3121 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3181 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgcatgca taaaaactgt
     3361 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     3421 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat tcaaatatgt
     3481 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3541 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3601 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3661 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3721 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3781 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3841 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3901 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3961 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4021 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4081 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4141 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4201 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4261 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4321 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4381 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4441 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4501 ccaaaatcca ggtggcactt ttcggggaaa tgtgcgcgcc cgcgttcctg ctggcgctgg
     4561 gcctgtttct ggcgctggac ttcccgctgt tccgtcagca gcttttcgcc cacggccttg
     4621 atgatcgcgg cggccttggc ctgcatatcc cgattcaacg gccccagggc gtccagaacg
     4681 ggcttcaggc gctcccgaag gtctcgggcc gtctcttggg cttgatcggc cttcttgcgc
     4741 atctcacgcg ctcctgcggc ggcctgtagg gcaggctcat acccctgccg aaccgctttt
     4801 gtcagccggt cggccacggc ttccggcgtc tcaacgcgct ttgagattcc cagcttttcg
     4861 gccaatccct gcggtgcata ggcgcgtggc tcgaccgctt gcgggctgat ggtgacgtgg
     4921 cccactggtg gccgctccag ggcctcgtag aacgcctgaa tgcgcgtgtg acgtgccttg
     4981 ctgccctcga tgccccgttg cagccctaga tcggccacag cggccgcaaa cgtggtctgg
     5041 tcgcgggtca tctgcgcttt gttgccgatg aactccttgg ccgacagcct gccgtcctgc
     5101 gtcagcggca ccacgaacgc ggtcatgtgc gggctggttt cgtcacggtg gatgctggcc
     5161 gtcacgatgc gatccgcccc gtacttgtcc gccagccact tgtgcgcctt ctcgaagaac
     5221 gccgcctgct gttcttggct ggccgacttc caccattccg ggctggccgt catgacgtac
     5281 tcgaccgcca acacagcgtc cttgcgccgc ttctctggca gcaactcgcg cagtcggccc
     5341 atcgcttcat cggtgctgct ggccgcccag tgctcgttct ctggcgtcct gctggcgtca
     5401 gcgttgggcg tctcgcgctc gcggtaggcg tgcttgagac tggccgccac gttgcccatt
     5461 ttcgccagct tcttgcatcg catgatcgcg tatgccgcca tgcctgcccc tcccttttgg
     5521 tgtccaaccg gctcgacggg ggcagcgcaa ggcggtgcct ccggcgggcc actcaatgct
     5581 tgagtatact cactagactt tgcttcgcaa agtcgtgacc gcctacggcg gctgcggcgc
     5641 cctacgggct tgctctccg
//
