LOCUS       sequence-313272-        7005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, pBBR origin, NahR(AM) repressor,
            PSalTTC promoter, GFPmut3b, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7005)
  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 7005)
  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: pBNG2
FEATURES             Location/Qualifiers
     source          1..7005
                     /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        3343..3392
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3437..3448
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3455..4168
                     /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(3485..3513)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4108..4128
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    4175..4195
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4196..4253
                     /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(4277..4294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(4732..5526)
                     /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(4843..4862)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5453..5472
                     /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 gattatcaat ccgtaaacag gtcaaacatc agttgccgca accaaatatt ggctaggtcc
     2221 ttgtggtact tcgcatgcca gaacatgttg atggctattt caggcaagac gactgggtgc
     2281 ggcaaggcgc ttaggccgaa gggctctacg cagcagtcgg ctaaacatat cggcacagtg
     2341 gcgagcagat cggtgcgctg gaggatgtgg ccaacggcgg cgaagtgcgg cacttccaga
     2401 cggatgtcgc gccggatgcc gacccgtgtc atgtacgtgt ccacctcgcc gtggccggtg
     2461 ccagcggcga tgacacgcac gtggccgtag gaacagaagc gctccagagt caggggttcg
     2521 cgggtgactg gatggtcctt gcgacatagg cacacgtagt gattctggag cagccggcgc
     2581 tgaaagaagc cagtttgcag attgggaagc aggcccacgg ccaagtccac ggttccgttc
     2641 tgcaaggcct gcatcaggct catcgaactg tcgcgcaccg tactgatcac gcaattgggg
     2701 gcctggtgag ccagcgcatc catcagccgc ggcatgaagt agatctcgcc aatgtcggtc
     2761 atggccaggg tgaaggtacg ctcgctggtc agcggatcga agctttcatg gtgctgtagg
     2821 gcgttgcgca gtgcgtgcat ggccgaagtg acgtgctcgg ccagatgcgc ggcatagggt
     2881 gtgggttcca ttccctgatg tgtgcgcacg aagagtgggt cctgtagcga ggtgcgcagg
     2941 cgtttcagcg cattgctcac ggcaggctgg gtcaggccca ggttctccgc agtgacagag
     3001 acgcgtctgt cgaccagcaa ctggttgaac accaccagca ggtttaaatc aaggtcacgc
     3061 agttccatgg atagttaagt ctttttctta tagggggtgg gcgctatcat gccataccgc
     3121 gaaaggtttt gcgccattcg atggcgcgcc gcccaattat tgaaggcctc cctaacgggg
     3181 ggcctttttt tgtttctggt ctcccgctta acgatcgttg gctgggggcc tcgcttgggt
     3241 tattgctggt gcccggccgg gcgcaatatt catgttgatg atttattata tatcgagtgg
     3301 tgtatttatt tatattgttt gctccgttac cgttattaac agctgtcacc ggatgtgctt
     3361 tccggtctga tgagtccgtg aggacgaaac agcctctaca aataattttg tttaagaact
     3421 gggaagagcg cagattaaag aggagaaata ctagatgcgt aaaggagaag aacttttcac
     3481 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
     3541 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
     3601 tactggaaaa ctacctgttc catggccaac acttgtcact actttcggtt atggtgttca
     3661 atgctttgcg agatacccag atcatatgaa acagcatgac tttttcaaga gtgccatgcc
     3721 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
     3781 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     3841 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
     3901 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
     3961 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
     4021 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     4081 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     4141 tacacatggc atggatgaac tatacaaata ataatactag tagcggccgc tgcagtccgg
     4201 caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc
     4261 gcgttatgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4321 cggtatcagc tcactcaaag gcggtaatgc tttccagtcg ggaaacctgt cgtgccagct
     4381 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gcatgcataa
     4441 aaactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     4501 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggg
     4561 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     4621 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     4681 atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa
     4741 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     4801 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     4861 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     4921 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     4981 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     5041 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     5101 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     5161 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     5221 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     5281 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     5341 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     5401 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     5461 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     5521 aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca
     5581 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     5641 gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta
     5701 tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt
     5761 ccagatagcc cagtagctga cattcatccc aggtggcact tttcgtggaa atgtgcgcgc
     5821 ccgcgttcct gctggcgctg ggcctgtttc tggcgctgga cttcccgctg ttccgtcagc
     5881 agcttttcgc ccacggcctt gatgatcgcg gcggccttgg cctgcatatc ccgattcaac
     5941 ggccccaggg cgtccagaac ggtcttcagg cgctcccgaa ggtctcgggc cgtctcttgg
     6001 gcttgatcgg ccttcttgcg catctcacgc gctcctgcgg cggcctgtag ggcaggctca
     6061 tacccctgcc gaaccgcttt tgtcagccgg tcggccacgg cttccggcgt ctcaacgcgc
     6121 tttgagattc ccagcttttc ggccaatccc tgcggtgcat aggcgcgtgg ctcgaccgct
     6181 tgcgggctga tggtgacgtg gcccactggt ggccgctcca gggcctcgta gaacgcctga
     6241 atgcgcgtgt gacgtgcctt gctgccctcg atgccccgtt gcagccctag atcggccaca
     6301 gcggccgcaa acgtggtctg gtcgcgggtc atctgcgctt tgttgccgat gaactccttg
     6361 gccgacagcc tgccgtcctg cgtcagcggc accacgaacg cggtcatgtg cgggctggtt
     6421 tcgtcacggt ggatgctggc cgtcacgatg cgatccgccc cgtacttgtc cgccagccac
     6481 ttgtgcgcct tctcgaagaa cgccgcctgc tgttcttggc tggccgactt ccaccattcc
     6541 gggctggccg tcatgacgta ctcgaccgcc aacacagcgt ccttgcgccg cttctctggc
     6601 agcaactcgc gcagtcggcc catcgcttca tcggtgctgc tggccgccca gtgctcgttc
     6661 tctggcgtcc tgctggcgtc agcgttgggc gtctcgcgct cgcggtaggc gtgcttgaga
     6721 ctggccgcca cgttgcccat tttcgccagc ttcttgcatc gcatgatcgc gtatgccgcc
     6781 atgcctgccc ctcccttttg gtgtccaacc ggctcgacgg gggcagcgca aggcggtgcc
     6841 tccggcgggc cactcaatgc ttgagtatac tcactagact ttgcttcgca aagtagtgac
     6901 cgcctacggc ggctgcggcg ccctacgggc ttgctctcag ggcttcgccc tgcgcggtcg
     6961 ctgcgctccc ttgccagccc gtggatatgt ggacgatggc cgcga
//
