LOCUS       sequence-206296-        3849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive LlacO-1 promoter expressing gfpuv on BBR1 origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3849)
  AUTHORS   Cook TB, Rand JM, Nurani W, Courtney DK, Liu SA, Pfleger BF
  TITLE     Genetic tools for reliable gene expression and recombineering in 
            Pseudomonas putida.
  JOURNAL   J Ind Microbiol Biotechnol. 2018 Jan 3. pii: 
            10.1007/s10295-017-2001-5. doi: 10.1007/s10295-017-2001-5.
  PUBMED    29299733
REFERENCE   2  (bases 1 to 3849)
  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: "J Ind 
            Microbiol Biotechnol. 2018 Jan 3. pii: 10.1007/s10295-017-2001-5. 
            doi: 10.1007/s10295-017-2001-5."
COMMENT     Sequence Label: pBBR6k-con-GFPuv
FEATURES             Location/Qualifiers
     source          1..3849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     202..221
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    289..305
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    312..328
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             396..1112
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(426..454)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1049..1069
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1145..1216
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1232..1259
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1326..1988)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      1989..2760
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(2794..2888)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2919..3713)
                     /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(3030..3049)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3640..3659
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtctccg gttcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
       61 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
      121 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
      181 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
      241 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagaatt gtgagcggat
      301 aacaattgac attgtgagcg gataacaaga tactgagcac atcagcagga cgcactgacc
      361 gaattcaaaa gatcttttaa gaaggagata tacatatgag taaaggagaa gaacttttca
      421 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      481 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
      541 ctactggaaa actacctgtt ccgtggccaa cacttgtcac tactttctct tatggtgttc
      601 aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc
      661 ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggaac tacaagacgc
      721 gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aaaggtattg
      781 attttaaaga agatggaaac attctcggac acaaactgga gtacaactat aactcacaca
      841 atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc
      901 acaacattga agatggctcc gttcaactag cagaccatta tcaacaaaat actccaattg
      961 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     1021 aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga
     1081 ttacacatgg catggatgag ctctacaaat aaggatccaa actcgagtaa ggatctccag
     1141 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1201 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     1261 ttatacctag gctacagccg atagtctgga acagcgcact tacgggttgc tgcgcaaccc
     1321 aagtgctacc ggcgcggcag cgtgacccgt gtcggcggct ccaacggctc gccatcgtcc
     1381 agaaaacacg gctcatcggg catcggcagg cgctgctgcc cgcgccgttc ccattcctcc
     1441 gtttcggtca aggctggcag gtctggttcc atgcccggaa tgccgggctg gctgggcggc
     1501 tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat acagggtcgg gatgcggcgc
     1561 aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac cacggcggca
     1621 ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg gtcattgacc
     1681 acgtaggccg acacggtgcc ggggccgttg agcttcacga cggagatcca gcgctcggcc
     1741 accaagtcct tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag cttggaaagt
     1801 gtcttctggc tgaccaccac ggcgttctgg tggcccatct gcgccacgag gtgatgcagc
     1861 agcattgccg ccgtgggttt cctcgcaata agcccggccc acgcctcatg cgctttgcgt
     1921 tccgtttgca cccagtgacc gggcttgttc ttggcttgaa tgccgatttc tctggactgc
     1981 gtggccatgc ttatctccat gcggtagggg tgccgcacgg ttgcggcacc atgcgcaatc
     2041 agctgcaact tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg gcagtcaatt
     2101 acagattttc tttaacctac gcaatgagct attgcggggg gtgccgcaat gagctgttgc
     2161 gtacccccct tttttaagtt gttgattttt aagtctttcg catttcgccc tatatctagt
     2221 tctttggtgc ccaaagaagg gcacccctgc ggggttcccc cacgccttcg gcgcggctcc
     2281 ccctccggca aaaagtggcc cctccggggc ttgttgatcg actgcgcggc cttcggcctt
     2341 gcccaaggtg gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg ctcggggggc
     2401 aggcgggcgg gcttcgccct tcgactgccc ccactcgcat aggcttgggt cgttccaggc
     2461 gcgtcaaggc caagccgctg cgcggtcgct gcgcgagtct tgacccgcct tccacttggt
     2521 gtccaaccgg caagcgaagc gcgcaggccg caggccggag gcttttcccc agagaaaatt
     2581 aaaaaaattg atggggcaag gccgcaggcc gcgcagttgg agccggtggg tatgtggtcg
     2641 aaggctgggt agccggtggg caatccctgt ggtcaagctc gtgggcaggc gcagcctgtc
     2701 catcagcttg tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc cggtggccgc
     2761 tcgcggccat cgtccacata tcctagtgct tggattctca ccaataaaaa acgcccggcg
     2821 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     2881 caggagtcca agcgagctct cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     2941 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     3001 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     3061 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     3121 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     3181 atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     3241 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     3301 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     3361 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     3421 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     3481 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca
     3541 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     3601 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     3661 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     3721 gatcctcatc ctgtctcttg atcagatcat gatcccctgc gccatcagat ccttggcggc
     3781 aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct
     3841 ggcaattcc
//
