LOCUS       sequence-394483-        8343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BHR plasmid (pBBR1 ori) expressing T7 polymerase under control of 
            the lac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8343)
  AUTHORS   Greenwich JL, Alakavuklar MA, Fuqua C
  TITLE     An Inducible T7 Polymerase System for High-Level Protein Expression 
            in Diverse Gram-Negative Bacteria.
  JOURNAL   Microbiol Resour Announc. 2023 Feb 16;12(2):e0111922. doi: 
            10.1128/mra.01119-22. Epub 2023 Jan 16.
  PUBMED    36645284
REFERENCE   2  (bases 1 to 8343)
  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: "2023-02-16"; volume: "12(2):e0111922. doi";
            pages: " 10.1128/mra.01119-22. Epub 2023 Jan 16"
COMMENT     Sequence Label: pJLG038
FEATURES             Location/Qualifiers
     source          1..8343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1427..1960)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2149..2154,2155..2171,2172..2177))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2421..2498
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2499..3581
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2518..2537)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    3594..3615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3630..3635,3636..3653,3654..3660)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3668..3684
                     /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)."
     primer_bind     3673..3695
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        3730..3748
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6477..6496)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6478..6496)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6503..6520)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6503..6519)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6512..6534)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(7054..7716)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      join(7717..8343,1..145)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 gttggagccg gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc cctgtggtca
       61 agctcgtggg caggcgcagc ctgtccatca gcttgtccag cagggttgtc cacgggccga
      121 gcgaagcgag ccagccggtg gccgctcgcg gccatcgtcc acatatccac gggctggcaa
      181 gggagcgcag cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg cgccgcagcc
      241 gccgtaggcg gtcacgactt tgcgaagcaa agtctagtga gtatactcaa gcattgagtg
      301 gcccgccgga ggcaccgcct tgcgctgccc ccgtcgagcc ggttggacac caaaagggag
      361 gggcaggcat ggcggcatac gcgatcatgc gatgcaagaa gctggcgaaa atgggcaacg
      421 tggcggccag tctcaagcac gcctaccgcg agcgcgagac gcccaacgct gacgccagca
      481 ggacgccaga gaacgagcac tgggcggcca gcagcaccga tgaagcgatg ggccgactgc
      541 gcgagttgct gccagagaag cggcgcaagg acgctgtgtt ggcggtcgag tacgtcatga
      601 cggccagccc ggaatggtgg aagtcggcca gccaagaaca gcaggcggcg ttcttcgaga
      661 aggcgcacaa gtggctggcg gacaagtacg gggcggatcg catcgtgacg gccagcatcc
      721 accgtgacga aaccagcccg cacatgaccg cgttcgtggt gccgctgacg caggacggca
      781 ggctgtcggc caaggagttc atcggcaaca aagcgcagat gacccgcgac cagaccacgt
      841 ttgcggccgc tgtggccgat ctagggctgc aacggggcat cgagggcagc aaggcacgtc
      901 acacgcgcat tcaggcgttc tacgaggccc tggagcggcc accagtgggc cacgtcacca
      961 tcagcccgca agcggtcgag ccacgcgcct atgcaccgca gggattggcc gaaaagctgg
     1021 gaatctcaaa gcgcgttgag acgccggaag ccgtggccga ccggctgaca aaagcggttc
     1081 ggcaggggta tgagcctgcc ctacaggccg ccgcaggagc gcgtgagatg cgcaagaagg
     1141 ccgatcaagc ccaagagacg gcccgagacc ttcgggagcg cctgaagccc gttctggacg
     1201 ccctggggcc gttgaatcgg gatatgcagg ccaaggccgc cgcgatcatc aaggccgtgg
     1261 gcgaaaagct gctgacggaa cagcgggaag tccagcgcca gaaacaggcc cagcgccagc
     1321 aggaacgcgg gcgcgcacat ttccccgaaa agtgccacct gcggcgttgt gacaatttac
     1381 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     1441 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     1501 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     1561 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctctccgg
     1621 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     1681 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     1741 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     1801 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     1861 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     1921 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     1981 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     2041 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     2101 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     2161 gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtccatgg
     2221 gcaaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2281 taggccgtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
     2341 cccttaaacg cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     2401 ttcgaaattg aattctgatt gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
     2461 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     2521 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     2581 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     2641 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     2701 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     2761 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     2821 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     2881 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     2941 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     3001 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     3061 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     3121 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     3181 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     3241 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     3301 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     3361 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     3421 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     3481 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3541 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3601 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3661 tgtggaattg tgagcggata acaatttcac acaggaaaca gcatatgacc atgattacgc
     3721 caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg
     3781 ccgctctaga atgaacacga ttaacatcgc taagaacgac ttctctgaca tcgaactggc
     3841 tgctatcccg ttcaacactc tggctgacca ttacggtgag cgtttagctc gcgaacagtt
     3901 ggcccttgag catgagtctt acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg
     3961 tcaacttaaa gctggtgagg ttgcggataa cgctgccgcc aagcctctca tcactaccct
     4021 actccctaag atgattgcac gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg
     4081 caagcgcccg acagccttcc agttcctgca agaaatcaag ccggaagccg tagcgtacat
     4141 caccattaag accactctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt
     4201 agcaagcgca atcggtcggg ccattgagga cgaggctcgc ttcggtcgta tccgtgacct
     4261 tgaagctaag cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt
     4321 ctacaagaaa gcatttatgc aagttgtcga ggctgacatg ctctctaagg gtctactcgg
     4381 tggcgaggcg tggtcttcgt ggcataagga agactctatt catgtaggag tacgctgcat
     4441 cgagatgctc attgagtcaa ccggaatggt tagcttacac cgccaaaatg ctggcgtagt
     4501 aggtcaagac tctgagacta tcgaactcgc acctgaatac gctgaggcta tcgcaacccg
     4561 tgcaggtgcg ctggctggca tctctccgat gttccaacct tgcgtagttc ctcctaagcc
     4621 gtggactggc attactggtg gtggctattg ggctaacggt cgtcgtcctc tggcgctggt
     4681 gcgtactcac agtaagaaag cactgatgcg ctacgaagac gtttacatgc ctgaggtgta
     4741 caaagcgatt aacattgcgc aaaacaccgc atggaaaatc aacaagaaag tcctagcggt
     4801 cgccaacgta atcaccaagt ggaagcattg tccggtcgag gacatccctg cgattgagcg
     4861 tgaagaactc ccgatgaaac cggaagacat cgacatgaat cctgaggctc tcaccgcgtg
     4921 gaaacgtgct gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag
     4981 ccttgagttc atgcttgagc aagccaataa gtttgctaac cataaggcca tctggttccc
     5041 ttacaacatg gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa
     5101 cgatatgacc aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta
     5161 ctactggctg aaaatccacg gtgcaaactg tgcgggtgtc gataaggttc cgttccctga
     5221 gcgcatcaag ttcattgagg aaaaccacga gaacatcatg gcttgcgcta agtctccact
     5281 ggagaacact tggtgggctg agcaagattc tccgttctgc ttccttgcgt tctgctttga
     5341 gtacgctggg gtacagcacc acggcctgag ctataactgc tcccttccgc tggcgtttga
     5401 cgggtcttgc tctggcatcc agcacttctc cgcgatgctc cgagatgagg taggtggtcg
     5461 cgcggttaac ttgcttccta gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa
     5521 agtcaacgag attctacaag cagacgcaat caatgggacc gataacgaag tagttaccgt
     5581 gaccgatgag aacactggtg aaatctctga gaaagtcaag ctgggcacta aggcactggc
     5641 tggtcaatgg ctggcttacg gtgttactcg cagtgtgact aagcgttcag tcatgacgct
     5701 ggcttacggg tccaaagagt tcggcttccg tcaacaagtg ctggaagata ccattcagcc
     5761 agctattgat tccggcaagg gtctgatgtt cactcagccg aatcaggctg ctggatacat
     5821 ggctaagctg atttgggaat ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa
     5881 ctggcttaag tctgctgcta agctgctggc tgctgaggtc aaagataaga agactggaga
     5941 gattcttcgc aagcgttgcg ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca
     6001 ggaatacaag aagcctattc agacgcgctt gaacctgatg ttcctcggtc agttccgctt
     6061 acagcctacc attaacacca acaaagatag cgagattgat gcacacaaac aggagtctgg
     6121 tatcgctcct aactttgtac acagccaaga cggtagccac cttcgtaaga ctgtagtgtg
     6181 ggcacacgag aagtacggaa tcgaatcttt tgcactgatt cacgactcct tcggtaccat
     6241 tccggctgac gctgcgaacc tgttcaaagc agtgcgcgaa actatggttg acacatatga
     6301 gtcttgtgat gtactggctg atttctacga ccagttcgct gaccagttgc acgagtctca
     6361 attggacaaa atgccagcac ttccggctaa aggtaacttg aacctccgtg acatcttaga
     6421 gtcggacttc gcgttcgcgt aagtcgacct cgaggggggg cccggtaccc aattcgccct
     6481 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     6541 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     6601 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     6661 aattgtaagc tagcaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg
     6721 ttaaatagcc gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt
     6781 gaccgtgtgc ttctcaaatg cctgaggcca gtttgctcag gctctccccg tggaggtaat
     6841 aattgacgat atgatcattt attctgcctc ccagagcctg ataaaaacgg tgaatccgtt
     6901 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     6961 aacgcgggga ggcagacaag gtatagggcg gcgaggcggc tacagccgat agtctggaac
     7021 agcgcactta cgggttgctg cgcaacccaa gtgctaccgg cgcggcagcg tgacccgtgt
     7081 cggcggctcc aacggctcgc catcgtccag aaaacacggc tcatcgggca tcggcaggcg
     7141 ctgctgcccg cgccgttccc attcctccgt ttcggtcaag gctggcaggt ctggttccat
     7201 gcccggaatg ccgggctggc tgggcggctc ctcgccgggg ccggtcggta gttgctgctc
     7261 gcccggatac agggtcggga tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg
     7321 gtcgtcgtga tcaaccacca cggcggcact gaacaccgac aggcgcaact ggtcgcgggg
     7381 ctggccccac gccacgcggt cattgaccac gtaggccgac acggtgccgg ggccgttgag
     7441 cttcacgacg gagatccagc gctcggccac caagtccttg actgcgtatt ggaccgtccg
     7501 caaagaacgt ccgatgagct tggaaagtgt cttctggctg accaccacgg cgttctggtg
     7561 gcccatctgc gccacgaggt gatgcagcag cattgccgcc gtgggtttcc tcgcaataag
     7621 cccggcccac gcctcatgcg ctttgcgttc cgtttgcacc cagtgaccgg gcttgttctt
     7681 ggcttgaatg ccgatttctc tggactgcgt ggccatgctt atctccatgc ggtaggggtg
     7741 ccgcacggtt gcggcaccat gcgcaatcag ctgcaacttt tcggcagcgc gacaacaatt
     7801 atgcgttgcg taaaagtggc agtcaattac agattttctt taacctacgc aatgagctat
     7861 tgcggggggt gccgcaatga gctgttgcgt accccccttt tttaagttgt tgatttttaa
     7921 gtctttcgca tttcgcccta tatctagttc tttggtgccc aaagaagggc acccctgcgg
     7981 ggttccccca cgccttcggc gcggctcccc ctccggcaaa aagtggcccc tccggggctt
     8041 gttgatcgac tgcgcggcct tcggccttgc ccaaggtggc gctgccccct tggaaccccc
     8101 gcactcgccg ccgtgaggct cggggggcag gcgggcgggc ttcgcccttc gactgccccc
     8161 actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc
     8221 gcgagccttg acccgccttc cacttggtgt ccaaccggca agcgaagcgc gcaggccgca
     8281 ggccggaggc ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc
     8341 gca
//
