LOCUS       sequence-315040-        6865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, pBBR origin, LacI repressor, 
            PLlacO-1 promoter, AmpR, Gentamicin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6865)
  AUTHORS   Schuster LA, Reisch CR
  TITLE     A plasmid toolbox for controlled gene expression across the 
            Proteobacteria.
  JOURNAL   Nucleic Acids Res. 2021 Jun 14. pii: 6298615. doi: 
            10.1093/nar/gkab496.
  PUBMED    34125913
REFERENCE   2  (bases 1 to 6865)
  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: "Nucleic 
            Acids Res. 2021 Jun 14. pii: 6298615. doi: 10.1093/nar/gkab496."
COMMENT     Sequence Label: pBLlCa5
FEATURES             Location/Qualifiers
     source          1..6865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1272..2043
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2044..2706
                     /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     3400..3422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3414..3431
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3415..3431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3482..4564)
                     /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     4526..4545
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4565..4642)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      complement(4706..4749)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4752..4773
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    4776..4792
                     /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)."
     protein_bind    4799..4815
                     /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)."
     CDS             join(4911..4979,4980..5771)
                     /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(5129..5148)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        join(6105..6110,6111..6127,6128..6133)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6322..6855
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 aagccgagat cggcttcccg gccgcggagt tgttcggtaa attgtcacaa cgccgcaggt
       61 ggcacttttc ggggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc
      121 gctggacttc ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg
      181 ccttggcctg catatcccga ttcaacggcc ccagggcgtc cagaacgggc ttcaggcgct
      241 cccgaaggtc tcgggccgtc tcttgggctt gatcggcctt cttgcgcatc tcacgcgctc
      301 ctgcggcggc ctgtagggca ggctcatacc cctgccgaac cgcttttgtc agccggtcgg
      361 ccacggcttc cggcgtctca acgcgctttg agattcccag cttttcggcc aatccctgcg
      421 gtgcataggc gcgtggctcg accgcttgcg ggctgatggt gacgtggccc actggtggcc
      481 gctccagggc ctcgtagaac gcctgaatgc gcgtgtgacg tgccttgctg ccctcgatgc
      541 cccgttgcag ccctagatcg gccacagcgg ccgcaaacgt ggtctggtcg cgggtcatct
      601 gcgctttgtt gccgatgaac tccttggccg acagcctgcc gtcctgcgtc agcggcacca
      661 cgaacgcggt catgtgcggg ctggtttcgt cacggtggat gctggccgtc acgatgcgat
      721 ccgccccgta cttgtccgcc agccacttgt gcgccttctc gaagaacgcc gcctgctgtt
      781 cttggctggc cgacttccac cattccgggc tggccgtcat gacgtactcg accgccaaca
      841 cagcgtcctt gcgccgcttc tctggcagca actcgcgcag tcggcccatc gcttcatcgg
      901 tgctgctggc cgcccagtgc tcgttctctg gcgtcctgct ggcgtcagcg ttgggcgtct
      961 cgcgctcgcg gtaggcgtgc ttgagactgg ccgccacgtt gcccattttc gccagcttct
     1021 tgcatcgcat gatcgcgtat gccgccatgc ctgcccctcc cttttggtgt ccaaccggct
     1081 cgacgggggc agcgcaaggc ggtgcctccg gcgggccact caatgcttga gtatactcac
     1141 tagactttgc ttcgcaaagt cgtgaccgcc tacggcggct gcggcgccct acgggcttgc
     1201 tctccgggct tcgccctgcg cggtcgctgc gctcccttgc catcccgtgg atatgtggac
     1261 gatggccgcg agcggccacc ggctggctcg cttcgctcgg cccgtggaca accctgctgg
     1321 acaagctgat ggacaggctg cgcctgccca cgagcttgac cacagggatt gcccaccggc
     1381 tacccagcct tcgaccacat acccaccggc tccaactgcg cggcctgcgg ccttgcccca
     1441 tcaatttttt taattttctc tggggaaaag cctccggcct gcggcctgcg cgcttcgctt
     1501 gccggttgga caccaagtgg aaggcgggtc aaggctcgcg cagcgaccgc gcagcggctt
     1561 ggccttgacg cgcctggaac gacccaagcc tatgcgagtg ggggcagtcg aagggcgaag
     1621 cccgcccgcc tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg
     1681 ccaccttggg caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt
     1741 ttgccggagg gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg
     1801 ggcaccaaag aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa
     1861 aaggggggta cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa
     1921 agaaaatctg taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa
     1981 aagttgcagc tgattgcgca tggtgccgca accgtgcggc acccctaccg catggagata
     2041 agcatggcca cgcagtccag agaaatcggc attcaagcca agaacaagcc cggtcactgg
     2101 gtgcaaacgg aacgcaaagc gcatgaggcg tgggccgggc ttattgcgag gaaacccacg
     2161 gcggcaatgc tgctgcatca cctcgtggcg cagatgggcc accagaacgc cgtggtggtc
     2221 agccagaaga cactttccaa gctcatcgga cgttctttgc ggacggtcca atacgcagtc
     2281 aaggacttgg tggccgagcg ctggatctcc gtcgtgaagc tcaacggccc cggcaccgtg
     2341 tcggcctacg tggtcaatga ccgcgtggcg tggggccagc cccgcgacca gttgcgcctg
     2401 tcggtgttca gtgccgccgt ggtggttgat cacgacgacc aggacgaatc gctgttgggg
     2461 catggcgacc tgcgccgcat cccgaccctg tatccgggcg agcagcaact accgaccggc
     2521 cccggcgagg agccgcccag ccagcccggc attccgggca tggaaccaga cctgccagcc
     2581 ttgaccgaaa cggaggaatg ggaacggcgc gggcagcagc gcctgccgat gcccgatgag
     2641 ccgtgttttc tggacgatgg cgagccgttg gagccgccga cacgggtcac gctgccgcgc
     2701 cggtagcact tgggttgcgc agcaacccgt aagtgcgctg ttccagacta tcggctgtag
     2761 ccgcctcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     2821 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cgtttttatc
     2881 aggctctggg aggcagaata aatgatcata tcgtcaatta ttacctccac ggggagagcc
     2941 tgagcaaact ggcctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
     3001 aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg
     3061 accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
     3121 cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     3181 gccactcatc gcagtacggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3241 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
     3301 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     3361 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     3421 cgacggccag tgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     3481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     3601 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     3661 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     3721 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     3781 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     3841 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     3901 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     3961 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     4021 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     4081 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     4141 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     4201 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     4261 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     4321 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     4381 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     4441 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     4501 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     4561 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     4621 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     4681 aggaaatggc gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag
     4741 gatgatttct ggaattcgcg gccgcttcta gagaattgtg agcggataac aattgacatt
     4801 gtgagcggat aacaagatac tgagcacacc ctgtaggaac tgggaagagc gcaccgctca
     4861 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4921 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4981 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5041 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5101 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5161 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5221 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5281 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5341 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5401 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5461 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5521 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5581 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5641 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5701 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5761 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5821 atttttaatt taaaaggatc taggtgaaga tcctttgctt tccagtcggg aaacctgtcg
     5881 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5941 atgcataaaa actgttgtaa ttcattaagc attctgccga catggaatcc atcacaaacg
     6001 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     6061 cccatggacg cacaccgtgg aaacggatga aggcacgaac ccagttgaca taagcctgtt
     6121 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     6181 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     6241 ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     6301 tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt
     6361 cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc
     6421 cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac
     6481 gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt
     6541 aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct
     6601 tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc
     6661 tccggagagc accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag
     6721 gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca
     6781 gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca
     6841 agtaccgcca cctaacaatt cgttc
//
