LOCUS       sequence-313649-        6666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, pBBR origin, TtgR(AM) repressor,
            PTtg promoter, GFPmut3b, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6666)
  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 6666)
  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: pBTtG2
FEATURES             Location/Qualifiers
     source          1..6666
                     /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        3004..3053
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3098..3109
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3116..3829
                     /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
                     NFKIRQNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3146..3174)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3769..3789
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    3836..3856
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3857..3914
                     /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(3938..3955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(4393..5187)
                     /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(4504..4523)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5114..5133
                     /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 gatcattatt tacgcagtgc cggactcaga cgcagcatat ccagaccggt atcaacccat
     2221 ttttcaacat cacccagcag atcaacgcta tccggcagca gcagccaacg acgaatcaga
     2281 ccatcaacat aggcaaacat tgcaactgcg gcacgttctg catccagttc accaggcagc
     2341 tgaccacgac gaactacatt tgccagtgtc agggtaatac ctttatgaca atccagaact
     2401 gcgctctgat gctgctgacg aatttcacac atatcatcgg tgaactcgca tttgtgatgc
     2461 aggatttcat taatacgacg ggtacgtgca tccagaacca gttcattaaa aacctgcagc
     2521 agcagtttac gcatacaacc cagcggatca acttcatctt cgctttcgct tgcacgtgcc
     2581 agatgatcat gggtttcatg caggctatcc agcagtgcct gaaccagttc ggctttgttg
     2641 ttaaaatgcc aataaattgc accacgggta acgcctgcca gttctgcaat atctgccagg
     2701 gtggtacgtg caacaccacg tttatagaat gcacgttctg ctgcttcaat aatctgtgca
     2761 cgggtttctt gtgcctcttc tttggtacga cgaaccatgt tttacctact tatctaattc
     2821 gtaagggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg cgcgccgccc
     2881 aattattgaa ggcctcccta acggggggcc tttttttgtt tctggtctcc cgcttaacga
     2941 tcgttggctg cacccagcag tatttacaaa caaccatgaa tgtaagtata ttccttagca
     3001 aagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac
     3061 aaataatttt gtttaagaac tgggaagagc gcagattaaa gaggagaaat actagatgcg
     3121 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
     3181 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
     3241 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
     3301 tactttcggt tatggtgttc aatgctttgc gagataccca gatcatatga aacagcatga
     3361 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
     3421 tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag
     3481 aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga
     3541 atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa
     3601 agttaacttc aaaattagac aaaacattga agatggaagc gttcaactag cagaccatta
     3661 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     3721 cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
     3781 gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaat aataatacta
     3841 gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt
     3901 taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct
     3961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaatg ctttccagtc
     4021 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4081 gcgtattggg cgcatgcata aaaactgttg taattcatta agcattctgc cgacatggaa
     4141 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     4201 cgtataatat ttgcccatgg gggtgggcga agaactccag catgagatcc ccgcgctgga
     4261 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     4321 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc
     4381 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     4441 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     4501 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     4561 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     4621 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag
     4681 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     4741 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     4801 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     4861 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     4921 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     4981 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt
     5041 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     5101 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     5161 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga
     5221 tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca
     5281 gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca
     5341 taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt
     5401 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc caggtggcac
     5461 ttttcgtgga aatgtgcgcg cccgcgttcc tgctggcgct gggcctgttt ctggcgctgg
     5521 acttcccgct gttccgtcag cagcttttcg cccacggcct tgatgatcgc ggcggccttg
     5581 gcctgcatat cccgattcaa cggccccagg gcgtccagaa cggtcttcag gcgctcccga
     5641 aggtctcggg ccgtctcttg ggcttgatcg gccttcttgc gcatctcacg cgctcctgcg
     5701 gcggcctgta gggcaggctc atacccctgc cgaaccgctt ttgtcagccg gtcggccacg
     5761 gcttccggcg tctcaacgcg ctttgagatt cccagctttt cggccaatcc ctgcggtgca
     5821 taggcgcgtg gctcgaccgc ttgcgggctg atggtgacgt ggcccactgg tggccgctcc
     5881 agggcctcgt agaacgcctg aatgcgcgtg tgacgtgcct tgctgccctc gatgccccgt
     5941 tgcagcccta gatcggccac agcggccgca aacgtggtct ggtcgcgggt catctgcgct
     6001 ttgttgccga tgaactcctt ggccgacagc ctgccgtcct gcgtcagcgg caccacgaac
     6061 gcggtcatgt gcgggctggt ttcgtcacgg tggatgctgg ccgtcacgat gcgatccgcc
     6121 ccgtacttgt ccgccagcca cttgtgcgcc ttctcgaaga acgccgcctg ctgttcttgg
     6181 ctggccgact tccaccattc cgggctggcc gtcatgacgt actcgaccgc caacacagcg
     6241 tccttgcgcc gcttctctgg cagcaactcg cgcagtcggc ccatcgcttc atcggtgctg
     6301 ctggccgccc agtgctcgtt ctctggcgtc ctgctggcgt cagcgttggg cgtctcgcgc
     6361 tcgcggtagg cgtgcttgag actggccgcc acgttgccca ttttcgccag cttcttgcat
     6421 cgcatgatcg cgtatgccgc catgcctgcc cctccctttt ggtgtccaac cggctcgacg
     6481 ggggcagcgc aaggcggtgc ctccggcggg ccactcaatg cttgagtata ctcactagac
     6541 tttgcttcgc aaagtagtga ccgcctacgg cggctgcggc gccctacggg cttgctctca
     6601 gggcttcgcc ctgcgcggtc gctgcgctcc cttgccagcc cgtggatatg tggacgatgg
     6661 ccgcga
//
