LOCUS       sequence-312895-        6971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, pBBR origin, CinR(AM) activator,
            PCin promoter, GFPmut3b, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  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: pBCiG2
FEATURES             Location/Qualifiers
     source          1..6971
                     /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"
     misc_RNA        3309..3358
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3403..3414
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3421..4134
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3451..3479)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4074..4094
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    4141..4161
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4162..4219
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(4243..4260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(4698..5492)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4809..4828)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5419..5438
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
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 agggcgaagc 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 gactattacc aattacgtcg cgtcatgcgg atcctatagg gattgatgat gcgcaattga
     2221 acagcccgcg acgcggcggc cgagatcgtg gtgcagccga gcctgaagcg ggcggttttc
     2281 aggtaatcgc gtgtggtatg ctctgatatg cccaggatga ccgaaatatc cttgtaatcc
     2341 ttgccgaggg cggtccagtg cagacactcg atctcgcgcg gcgacaatgc cggcaccgga
     2401 tcgttttcgc catgcagctc atatacggcc ttgcggtgga tcagatgggc gatctcgatc
     2461 cactcattgc ggcagcggcg cacgagctcg gtccattcgt cggccggtat atgggcattc
     2521 agcgacagca gggcgcggcg ctgcgccttg tcggcgacgg ggatggagta gccattgtca
     2581 tcgatgccgt gtttctgggc gtcgaccagc atggcatagg cctccggcgt cggttcgacc
     2641 tcgctccagt cgaagggcag ctggcgttcg aagccctgct tgatgatcgg atcgaccttc
     2701 acatagcagt tgaggaggta acgggaaacc caggcatccg gataggtggt gcgcacgaag
     2761 ggcgaatcga tcttgctcgc gattgtctgg gcgagatggt aggtgacgaa atcgaggtta
     2821 tattccgcct ggagaatacg gatggcggca tccacgttgg ccgccgcttt gatctgttcg
     2881 aacgcggact cgaacttttc gctataggta ttctcaatca ttcgccgttc tccgttgacg
     2941 ggcgtctcgg acctgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     3001 cgcgccgccc aattattgaa ggcctcccta acggggggcc tttttttgtt tctggtctcc
     3061 cgcttaacga tcgttggctg ccctttgtgc gtccaaacgg acgcacggcg ctctaaagcg
     3121 ggtcgcgatc tttcagattc gctcctcgcg ctttcagtct ttgttttggc gcatgtcgtt
     3181 atcgcaaaac cgctgcacac ttttgcgcga catgctctga tccccctcat ctgggggggc
     3241 ctatctgagg gaatttccga tccggctcgc ctgaaccatt ctgctttcca cgaacttgaa
     3301 aacgctagct gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc
     3361 tctacaaata attttgttta agaactggga agagcgcaga ttaaagagga gaaatactag
     3421 atgcgtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     3481 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
     3541 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
     3601 gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca tatgaaacag
     3661 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     3721 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
     3781 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
     3841 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
     3901 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     3961 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     4021 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     4081 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaataataa
     4141 tactagtagc ggccgctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt
     4201 ttctttaaaa ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct
     4261 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatgctttc
     4321 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4381 ggtttgcgta ttgggcgcat gcataaaaac tgttgtaatt cattaagcat tctgccgaca
     4441 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     4501 ccttgcgtat aatatttgcc catgggggtg ggcgaagaac tccagcatga gatccccgcg
     4561 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     4621 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     4681 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     4741 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     4801 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     4861 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     4921 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     4981 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     5041 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     5101 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     5161 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5221 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     5281 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     5341 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5401 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5461 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     5521 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     5581 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct
     5641 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt
     5701 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catcccaggt
     5761 ggcacttttc gtggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc
     5821 gctggacttc ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg
     5881 ccttggcctg catatcccga ttcaacggcc ccagggcgtc cagaacggtc ttcaggcgct
     5941 cccgaaggtc tcgggccgtc tcttgggctt gatcggcctt cttgcgcatc tcacgcgctc
     6001 ctgcggcggc ctgtagggca ggctcatacc cctgccgaac cgcttttgtc agccggtcgg
     6061 ccacggcttc cggcgtctca acgcgctttg agattcccag cttttcggcc aatccctgcg
     6121 gtgcataggc gcgtggctcg accgcttgcg ggctgatggt gacgtggccc actggtggcc
     6181 gctccagggc ctcgtagaac gcctgaatgc gcgtgtgacg tgccttgctg ccctcgatgc
     6241 cccgttgcag ccctagatcg gccacagcgg ccgcaaacgt ggtctggtcg cgggtcatct
     6301 gcgctttgtt gccgatgaac tccttggccg acagcctgcc gtcctgcgtc agcggcacca
     6361 cgaacgcggt catgtgcggg ctggtttcgt cacggtggat gctggccgtc acgatgcgat
     6421 ccgccccgta cttgtccgcc agccacttgt gcgccttctc gaagaacgcc gcctgctgtt
     6481 cttggctggc cgacttccac cattccgggc tggccgtcat gacgtactcg accgccaaca
     6541 cagcgtcctt gcgccgcttc tctggcagca actcgcgcag tcggcccatc gcttcatcgg
     6601 tgctgctggc cgcccagtgc tcgttctctg gcgtcctgct ggcgtcagcg ttgggcgtct
     6661 cgcgctcgcg gtaggcgtgc ttgagactgg ccgccacgtt gcccattttc gccagcttct
     6721 tgcatcgcat gatcgcgtat gccgccatgc ctgcccctcc cttttggtgt ccaaccggct
     6781 cgacgggggc agcgcaaggc ggtgcctccg gcgggccact caatgcttga gtatactcac
     6841 tagactttgc ttcgcaaagt agtgaccgcc tacggcggct gcggcgccct acgggcttgc
     6901 tctcagggct tcgccctgcg cggtcgctgc gctcccttgc cagcccgtgg atatgtggac
     6961 gatggccgcg a
//
