LOCUS       sequence-206288-        4699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L-arabinose-inducible araBAD promoter expressing gfpuv on BBR1 
            origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4699)
  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 4699)
  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: pBBR8k-GFPuv
FEATURES             Location/Qualifiers
     source          1..4699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..885)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        912..1196
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1119..1138
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1246..1962
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1276..1304)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1899..1919
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1995..2066
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2082..2109
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2176..2838)
                     /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      2839..3610
                     /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(3644..3738)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3769..4563)
                     /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(3880..3899)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4490..4509
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
       61 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
      121 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
      181 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
      241 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
      301 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
      361 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
      421 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
      481 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
      541 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
      601 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
      661 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
      721 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
      781 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
      841 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
      901 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
      961 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
     1021 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
     1081 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
     1141 aagattagcg gattctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
     1201 gtttttttgg gaattcaaaa gatcttttaa gaaggagata tacatatgag taaaggagaa
     1261 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     1321 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     1381 tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac tactttctct
     1441 tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag
     1501 agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggaac
     1561 tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta
     1621 aaaggtattg attttaaaga agatggaaac attctcggac acaaactgga gtacaactat
     1681 aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc
     1741 aaaattcgcc acaacattga agatggctcc gttcaactag cagaccatta tcaacaaaat
     1801 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     1861 gccctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact
     1921 gctgctggga ttacacatgg catggatgag ctctacaaat aaggatccaa actcgagtaa
     1981 ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2041 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     2101 ctttctgcgt ttatacctag gctacagccg atagtctgga acagcgcact tacgggttgc
     2161 tgcgcaaccc aagtgctacc ggcgcggcag cgtgacccgt gtcggcggct ccaacggctc
     2221 gccatcgtcc agaaaacacg gctcatcggg catcggcagg cgctgctgcc cgcgccgttc
     2281 ccattcctcc gtttcggtca aggctggcag gtctggttcc atgcccggaa tgccgggctg
     2341 gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat acagggtcgg
     2401 gatgcggcgc aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac
     2461 cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg
     2521 gtcattgacc acgtaggccg acacggtgcc ggggccgttg agcttcacga cggagatcca
     2581 gcgctcggcc accaagtcct tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag
     2641 cttggaaagt gtcttctggc tgaccaccac ggcgttctgg tggcccatct gcgccacgag
     2701 gtgatgcagc agcattgccg ccgtgggttt cctcgcaata agcccggccc acgcctcatg
     2761 cgctttgcgt tccgtttgca cccagtgacc gggcttgttc ttggcttgaa tgccgatttc
     2821 tctggactgc gtggccatgc ttatctccat gcggtagggg tgccgcacgg ttgcggcacc
     2881 atgcgcaatc agctgcaact tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg
     2941 gcagtcaatt acagattttc tttaacctac gcaatgagct attgcggggg gtgccgcaat
     3001 gagctgttgc gtacccccct tttttaagtt gttgattttt aagtctttcg catttcgccc
     3061 tatatctagt tctttggtgc ccaaagaagg gcacccctgc ggggttcccc cacgccttcg
     3121 gcgcggctcc ccctccggca aaaagtggcc cctccggggc ttgttgatcg actgcgcggc
     3181 cttcggcctt gcccaaggtg gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg
     3241 ctcggggggc aggcgggcgg gcttcgccct tcgactgccc ccactcgcat aggcttgggt
     3301 cgttccaggc gcgtcaaggc caagccgctg cgcggtcgct gcgcgagcct tgacccgcct
     3361 tccacttggt gtccaaccgg caagcgaagc gcgcaggccg caggccggag gcttttcccc
     3421 agagaaaatt aaaaaaattg atggggcaag gccgcaggcc gcgcagttgg agccggtggg
     3481 tatgtggtcg aaggctgggt agccggtggg caatccctgt ggtcaagctc gtgggcaggc
     3541 gcagcctgtc catcagcttg tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc
     3601 cggtggccgc tcgcggccat cgtccacata tcctagtgct tggattctca ccaataaaaa
     3661 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     3721 gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctc agaagaactc
     3781 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     3841 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     3901 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     3961 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     4021 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     4081 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     4141 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     4201 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     4261 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     4321 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     4381 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     4441 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     4501 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     4561 catgcgaaac gatcctcatc ctgtctcttg atcagatcat gatcccctgc gccatcagat
     4621 ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
     4681 cgccccagct ggcaattcc
//
