LOCUS       sequence-313680-        6792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, pBBR origin, VanR(AM) repressor,
            PVanCC promoter, GFPmut3b, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6792)
  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 6792)
  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: pBVG2
FEATURES             Location/Qualifiers
     source          1..6792
                     /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        3130..3179
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3224..3235
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3242..3955
                     /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(3272..3300)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3895..3915
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    3962..3982
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3983..4040
                     /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(4064..4081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(4519..5313)
                     /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(4630..4649)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5240..5259
                     /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 gaaacagacg aagaatccat ggtcaatctg cacgaattga ccatgctgca cccagcggtg
     2221 cgcctgcgct tgctgctgct tcaaaaactt ttgcattacg aattgctgcc agtgcatgat
     2281 cacgcataat acgttctgca ccttcggcat caccacagct aactgcatcc agaactgcct
     2341 gatgctgacg atgtgctgcc agcagatgtt catattcggc agacaggtcc atcagatcca
     2401 gggccagtgc accggctgct gcaaacggtt caaaaccatt acgtgccagt gcgctttcaa
     2461 ctgcaccatt acctgctgcg ctaaccaggg tatcatgaaa tgcctgatta tatgcggcat
     2521 aacgatccag atcttcacca ttcaggcgac cggctgcaaa cagtgcttca ccttctgcaa
     2581 tcagtacaac aaaacgtgca tgggtttctg aggtcatacc acgttctgcc agacgacgtg
     2641 ctgcaaaacc ttccagaaca ccacgaactt caattgcatc acgaatctga tcgctgctaa
     2701 caccacgggc tgcataacca cgtgcaccca gacgaacaac cagaccttct tgttccagtg
     2761 aacgcagtgc gatacgaacc ggcatacggc taacacccag tgctgctgcg gtcggaattt
     2821 ctgcaatacg ttcaccactt ttgatttcac cgcttgcaat cattttacgc agtgccatca
     2881 taacacgctg acccggttta atacgaggca tgtccatgtc tatcctcctt atttacgttc
     2941 gttagtttaa gcgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggcg
     3001 cgccgccaat tattgaaggc ctccctaacg gggggccttt ttttgtttct ggtctcccgc
     3061 ttaacgatcg ttggctgatt ggatccaatt gacagctagc tcagtcctag gtaccattgg
     3121 atccaatagc tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc
     3181 ctctacaaat aattttgttt aagaactggg aagagcgcag attaaagagg agaaatacta
     3241 gatgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     3301 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     3361 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     3421 tgtcactact ttcggttatg gtgttcaatg ctttgcgaga tacccagatc atatgaaaca
     3481 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
     3541 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
     3601 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     3661 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     3721 aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga
     3781 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     3841 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     3901 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaataata
     3961 atactagtag cggccgctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt
     4021 tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc
     4081 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatgcttt
     4141 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4201 cggtttgcgt attgggcgca tgcataaaaa ctgttgtaat tcattaagca ttctgccgac
     4261 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     4321 gccttgcgta taatatttgc ccatgggggt gggcgaagaa ctccagcatg agatccccgc
     4381 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     4441 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     4501 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     4561 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     4621 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     4681 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     4741 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     4801 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4861 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4921 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4981 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     5041 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     5101 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     5161 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     5221 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     5281 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     5341 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     5401 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
     5461 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
     5521 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatcccagg
     5581 tggcactttt cggggaaatg tgcgcgcccg cgttcctgct ggcgctgggc ctgtttctgg
     5641 cgctggactt cccgctgttc cgtcagcagc ttttcgccca cggccttgat gatcgcggcg
     5701 gccttggcct gcatatcccg attcaacggc cccagggcgt ccagaacggg cttcaggcgc
     5761 tcccaaaggt ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct
     5821 cctgcggcgg cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg
     5881 gccacggctt ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc
     5941 ggtgcatagg cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc
     6001 cgctccaggg cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg
     6061 ccccgttgca gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc
     6121 tgcgctttgt tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc
     6181 acgaacgcgg tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga
     6241 tccgccccgt acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt
     6301 tcttggctgg ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac
     6361 acagcgtcct tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg
     6421 gtgctgctgg ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc
     6481 tcgcgctcgc ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc
     6541 ttgcatcgca tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc
     6601 tcgacggggg cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca
     6661 ctagactttg cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg
     6721 ctctccgggc ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga
     6781 cgatggccgc ga
//
