LOCUS       sequence-313271-        6800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, pBBR origin, LuxR activator, 
            PLuxB promoter, GFPmut3b, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6800)
  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 6800)
  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: pBLxG2
FEATURES             Location/Qualifiers
     source          1..6800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1248..2019
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2020..2682
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETDEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     3376..3398
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3390..3407
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3391..3407
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        4385..4434
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             4479..4490
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4497..5210
                     /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(4527..4555)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5150..5170
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    5217..5237
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5238..5295
                     /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(5319..5336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(5774..6568)
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTCVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5885..5904)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6495..6514
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gtccagatag cccagtagct gacattcatc ccaggtggca cttttcgggg aaatgtgcgc
       61 gcccgcgttc ctgctggcgc tgggcctgtt tctggcgctg gacttcccgc tgttccgtca
      121 gcagcttttc gcccacggcc ttgatgatcg cggcggcctt ggcctgcata tcccgattca
      181 acggccccag ggcgtccaga acgggcttca ggcgctcccg aaggtctcgg gccgtctctt
      241 gggcttgatc ggccttcttg cgcatctcac gcgctcctgc ggcggcctgt agggcaggct
      301 catacccctg ccgaaccgct tttgtcagcc ggttggccac ggcttccggc gtctcaacgc
      361 gctttgagat tcccagcttt tcggccaatc cctgcggtgc ataggcgcgt ggctcgaccg
      421 cttgcgggct gatggtgacg tggcccactg gtggccgctc cagggcctcg tagaacgcct
      481 gaatgcgcgt gtgacgtgcc ttgctgccct cgatgccccg ttgcagccct agatcggcca
      541 cagcggccgc aaacgtggtc tggtcgcggg tcatctgcgc tttgttgccg atgaactcct
      601 tggccgacag cctgccgtcc tgcgtcagcg gcaccacgaa cgcggtcatg tgcgggctgg
      661 tttcgtcacg gtggatgctg gccgtcacga tgcgatccgc cccgtacttg tccgccagcc
      721 acttgtgcgc cttctcgaag aacgccgcct gctgttcttg gctggccgac ttccaccatt
      781 ccgggctggc cgtcatgacg tactcgaccg ccaacacagc gtccttgcgc cgcttctctg
      841 gcagcaactc gcgcagtcgg cccatcgctt catcggtgct gctggccgcc cagtgctcgt
      901 tctctggcgt cctgctggcg tcagcgttgg gcgtctcgcg ctcgcggtag gcgtgcttga
      961 gactggccgc cacgttgccc attttcgcca gcttcttgca tcgcatgatc gcgtatgccg
     1021 ccatgcctgc ccctcccttt tggtgtccaa ccggctcgac gggggcagcg caaggcggtg
     1081 cctccggcgg gccactcaat gcttgagtat actcactaga ctttgcttcg caaagtcgtg
     1141 accgcctacg gcggctgcgg cgccctacgg gcttgctctc cgggcttcgc cctgcgcggt
     1201 cgctgcgctc ccttgccagc ccgtggatat gtggacgatg gccgcgagcg gccaccggct
     1261 ggctcgcttc gctcggcccg tggacaaccc tgctggacaa gctgatggac aggctgcgcc
     1321 tgcccacgag cttgaccaca gggattgccc accggctacc cagccttcga ccacataccc
     1381 accggctcca actgcgcggc ctgcggcctt gccccatcaa tttttttaat tttctctggg
     1441 gaaaagcctc cggcctgcgg cctgcgcgct tcgcttgccg gttggacacc aagtggaagg
     1501 cgggtcaagg ctcgcgcagc gaccgcgcag cggcttggcc ttgacgcgcc tggaacgacc
     1561 caagcctatg cgagtggggg cagtcgaagg gcgaagcccg cccgcctgcc ccccgagcct
     1621 cacggcggcg agtgcggggg ttccaagggg gcagcgccac cttgggcaag gccgaaggcc
     1681 gcgcagtcga tcaacaagcc ccggaggggc cactttttgc cggaggggga gccgcgccga
     1741 aggcgtgggg gaaccccgca ggggtgccct tctttgggca ccaaagaact agatataggg
     1801 cgaaatgcga aagacttaaa aatcaacaac ttaaaaaagg ggggtacgca acagctcatt
     1861 gcggcacccc ccgcaatagc tcattgcgta ggttaaagaa aatctgtaat tgactgccac
     1921 ttttacgcaa cgcataattg ttgtcgcgct gccgaaaagt tgcagctgat tgcgcatggt
     1981 gccgcaaccg tgcggcaccc ctaccgcatg gagataagca tggccacgca gtccagagaa
     2041 atcggcattc aagccaagaa caagcccggt cactgggtgc aaacggaacg caaagcgcat
     2101 gaggcgtggg ccgggcttat tgcgaggaaa cccacggcgg caatgctgct gcatcacctc
     2161 gtggcgcaga tgggccacca gaacgccgtg gtggtcagcc agaagacact ttccaagctc
     2221 atcggacgtt ctttgcggac ggtccaatac gcagtcaagg acttggtggc cgagcgctgg
     2281 atctccgtcg tgaagctcaa cggccccggc accgtgtcgg cctacgtggt caatgaccgc
     2341 gtggcgtggg gccagccccg cgaccagttg cgcctgtctg tgttcagtgc cgccgtggtg
     2401 gttgatcacg acgaccagga cgaatcgctg ttggggcatg gcgacctgcg ccgcatcccg
     2461 accctgtatc cgggcgagca gcaactaccg accggccccg gcgaggagcc gcccagccag
     2521 cccggcattc cgggcatgga accagacctg ccagccttga ccgaaacgga cgaatgggaa
     2581 cggcgcgggc agcagcgcct gccgatgccc gatgagccgt gttttctgga cgatggcgag
     2641 ccgttggagc cgccgacacg ggtcacgctg ccgcgccggt agcacttggg ttgcgcagca
     2701 acccgtaagt gcgctgttcc agactatcgg ctgtagccgc ctcgccgccc tataccttgt
     2761 ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc
     2821 cggcggcacc tcgctaacgg attcaccgtt tttatcaggc tctgggaggc agaataaatg
     2881 atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc tcaggcattt
     2941 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca
     3001 taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat
     3061 ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc
     3121 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tacggcctat
     3181 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3241 cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3301 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3361 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgat caatttttaa
     3421 agtatgggca atcaattgct cctgttaaaa ttgctttaga aatactttgg cagcggtttg
     3481 ttgtattgag tttcatttgc gcattggtta aatggaaagt gacagtacgc tcactgcaac
     3541 ctaatatttt tgaaatatcc caagagcttt ttccttcgca tgcccacgct aaacattctt
     3601 tttctctttt ggttaaatcg ttgtttgatt tattatttgc tatatttatt tttcgataat
     3661 tatcaactag agaaggaaca attaatggta tgttcataca cgcatgtaaa aataaactat
     3721 ctatatagtt gtctttttct gaatgtgcaa aactaagcat tccgaagcca ttgttagccg
     3781 tatgaatagg gaaactaaac ccagtgataa gacctgatgt tttcgcttct ttaattacat
     3841 ttggagattt tttatttaca gcattgtttt caaatatatt ccaattaatt ggtgaatgat
     3901 tggagttaga ataatctact ataggatcat attttatcaa attagcgtca tcataatatt
     3961 gcctccattt tttagggtaa ttatctagga ttgaaatatc agatttaacc atagaatgag
     4021 gataaatgat cgcgagtaaa taatattcac aatgtaccat tttagtcata tcagataagc
     4081 attgattaat atcattattg cttctacaag ctttaatttt attaattatt ctgtatgtgt
     4141 cgtcggcatt tatgtttttc attctgcccc tccgaaaggc ttaagtcctc gcgtagtacc
     4201 tgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggcgc gccgcccaat
     4261 tattgaaggc ctccctaacg gggggccttt ttttgtttct ggtctcccgc ttaacgatcg
     4321 ttggctgacc tgtaggatcg tacaggttta cgcaagaaaa tggtttgtta cagtcgaata
     4381 aaagctgtca ccggatgtgc tttccggtct gatgagtccg tgaggacgaa acagcctcta
     4441 caaataattt tgtttaagaa ctgggaagag cgcagattaa agaggagaaa tactagatgc
     4501 gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     4561 ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac
     4621 ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca acacttgtca
     4681 ctactttcgg ttatggtgtt caatgctttg cgagataccc agatcatatg aaacagcatg
     4741 actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag
     4801 atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata
     4861 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
     4921 aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca
     4981 aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt
     5041 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     5101 ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg
     5161 agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa taataatact
     5221 agtagcggcc gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct
     5281 ttaaaaccga aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc
     5341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat gctttccagt
     5401 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5461 tgcgtattgg gcgcatgcat aaaaactgtt gtaattcatt aagcattctg ccgacatgga
     5521 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     5581 gcgtataata tttgcccatg ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     5641 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     5701 gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac
     5761 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     5821 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     5881 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     5941 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     6001 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     6061 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     6121 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     6181 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     6241 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     6301 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgcac gtcgtggcca
     6361 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     6421 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     6481 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     6541 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     6601 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     6661 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc
     6721 ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct
     6781 ttgcgcttgc gttttccctt
//
