LOCUS       sequence-247918-        7309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host-range plasmid with toxin-antitoxin system for maintenance
            in the absence of antibiotic selection, and constitutive promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7309)
  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 7309)
  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: pBBR5pemIKpKan
FEATURES             Location/Qualifiers
     source          1..7309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..772
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             773..1435
                     /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     2129..2151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2143..2160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2144..2160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2170..2189
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2170..2188
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2341..2357
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2341..2357
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2366..2490
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2515..2520,2521..2538,2539..2545)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2599..3258
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2665..2684)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3600..3905
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3784..3803
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3946..4070)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4809..4825)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4810..4826)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4855..4875)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4855..4873)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4894..4910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4894..4910)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4907..4929)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4918..4934
                     /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(4942..4948,4949..4966,4967..4972))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4987..5008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5278..5283,5284..5300,5301..5306)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5495..6028
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg
       61 gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt
      121 cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat caattttttt
      181 aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg ccggttggac
      241 accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg gccttgacgc
      301 gcctggaacg acccaagcct atgcgagtgg gggcagtcga aaggcgaagc ccgcccgcct
      361 gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc caccttgggc
      421 aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt tgccggaggg
      481 ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg gcaccaaaga
      541 actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa aggggggtac
      601 gcaacagctc attgcggcac cccccgcaat agctcattgc gtaggttaaa gaaaatctgt
      661 aattgactgc cacttttacg caacgcataa ttgttgtcgc gctgccgaaa agttgcagct
      721 gattgcgcat ggtgccgcaa ccgtgcggca cccctaccgc atggagataa gcatggccac
      781 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
      841 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
      901 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
      961 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     1021 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     1081 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     1141 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     1201 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     1261 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     1321 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     1381 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     1441 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     1501 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     1561 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     1621 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     1681 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     1741 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     1801 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca
     1861 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     1921 cagtacggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1981 caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg
     2041 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     2101 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     2161 gagcgcgcgt aatacgactc actatagggc gaattggagc tcccggaatt gccagctggg
     2221 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     2281 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc
     2341 tctagaacta gtggatcccc catcaacaag tttgtacaaa aaagctgaac gagaaacgta
     2401 aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata
     2461 ctgtaaaaca caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca
     2521 ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca
     2581 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     2641 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     2701 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     2761 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
     2821 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     2881 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     2941 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     3001 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     3061 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     3121 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     3181 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     3241 tggcagggcg gggcgtaaag atctggatcc ggcttactaa aagccagata acagtatgcg
     3301 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     3361 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     3421 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     3481 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     3541 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
     3601 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     3661 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     3721 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     3781 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     3841 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     3901 tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata
     3961 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
     4021 atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt tgatgggctg
     4081 caggaattcg attttcgcta taagcgtata aatggaggaa ttatattata ctttaatata
     4141 acttaacgaa agaagcccat catgcacacc acaaaccttc gtaaagtcgg tggatcgatc
     4201 atgctggccg ttccccccgc tttccttgat cagcttcatc ttgaggtcgg cgcaacggtc
     4261 ggcttagccg tcacagacgg tcgtctggtc attgagccag tcctacatcc tcagtacacg
     4321 ctggatcaac tgctggccga agcggaagcc tccggggctt acccgttgcc gcccgaggaa
     4381 cgtgaatggg tcgatgctcc agcagtagga cgcgaactga tatgaaacgc ggcgatgttt
     4441 acatggtcga tctggagcca accgccggac acgaacaacg cggtcaccgt ccggtggttg
     4501 ttatctccag tgagcgattc aatcggctta cctcttgtcc ggtgatcctt ccaatcacaa
     4561 acggcggcga gttcgcctcc cgcctcgggt tcgctgtgga attactaggt acgattacaa
     4621 cgggcgtcgt tcgatgtgat caaccacgtg cgctcgactt actcgctcgc aatgcccgca
     4681 aagtggaaag cctaccacca aacatcttgg ccgatgtctt ggcaaaagct gtcaccatct
     4741 tccagtaatt tttggggcaa gttcaggcca ctgcaaccaa tcactagtga attcgatatc
     4801 aagcttatcg ataccgtcga cctcgagggg gggcccggta cccagctttt gttcccttta
     4861 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4921 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4981 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5041 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5101 gcgtattggg cgcatgcata aaaactgttg taattcatta agcattctgc cgacatggaa
     5161 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     5221 cgtataatat ttgcccatgg acgcacaccg tggaaacgga tgaaggcacg aacccagttg
     5281 acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg
     5341 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     5401 tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     5461 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca acgatgttac
     5521 gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac
     5581 atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg
     5641 tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa
     5701 cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg
     5761 cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga tctatatcta
     5821 tgatctcgca gtctccggag agcaccggag gcagggcatt gccaccgcgc tcatcaatct
     5881 cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg
     5941 tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt
     6001 tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttcccggc
     6061 cgcggagttg ttcggtaaat tgtcacaacg ccgcaggtgg cacttttcgg ggaaatgtgc
     6121 gcgcccgcgt tcctgctggc gctgggcctg tttctggcgc tggacttccc gctgttccgt
     6181 cagcagcttt tcgcccacgg ccttgatgat cgcggcggcc ttggcctgca tatcccgatt
     6241 caacggcccc agggcgtcca gaacgggctt caggcgctcc cgaaggtctc gggccgtctc
     6301 ttgggcttga tcggccttct tgcgcatctc acgcgctcct gcggcggcct gtagggcagg
     6361 ctcatacccc tgccgaaccg cttttgtcag ccggtcggcc acggcttccg gcgtctcaac
     6421 gcgctttgag attcccagct tttcggccaa tccctgcggt gcataggcgc gtggctcgac
     6481 cgcttgcggg ctgatggtga cgtggcccac tggtggccgc tccagggcct cgtagaacgc
     6541 ctgaatgcgc gtgtgacgtg ccttgctgcc ctcgatgccc cgttgcagcc ctagatcggc
     6601 cacagcggcc gcaaacgtgg tctggtcgcg ggtcatctgc gctttgttgc cgatgaactc
     6661 cttggccgac agcctgccgt cctgcgtcag cggcaccacg aacgcggtca tgtgcgggct
     6721 ggtttcgtca cggtggatgc tggccgtcac gatgcgatcc gccccgtact tgtccgccag
     6781 ccacttgtgc gccttctcga agaacgccgc ctgctgttct tggctggccg acttccacca
     6841 ttccgggctg gccgtcatga cgtactcgac cgccaacaca gcgtccttgc gccgcttctc
     6901 tggcagcaac tcgcgcagtc ggcccatcgc ttcatcggtg ctgctggccg cccagtgctc
     6961 gttctctggc gtcctgctgg cgtcagcgtt gggcgtctcg cgctcgcggt aggcgtgctt
     7021 gagactggcc gccacgttgc ccattttcgc cagcttcttg catcgcatga tcgcgtatgc
     7081 cgccatgcct gcccctccct tttggtgtcc aaccggctcg acgggggcag cgcaaggcgg
     7141 tgcctccggc gggccactca atgcttgagt atactcacta gactttgctt cgcaaagtcg
     7201 tgaccgccta cggcggctgc ggcgccctac gggcttgctc tccgggcttc gccctgcgcg
     7261 gtcgctgcgc tcccttgcca gcccgtggat atgtggacga tggccgcga
//
