LOCUS       sequence-215245-        7505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescence microscopy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7505)
  AUTHORS   Lefebre MD, Valvano MA
  TITLE     Construction and evaluation of plasmid vectors optimized for 
            constitutive and regulated gene expression in Burkholderia cepacia 
            complex isolates.
  JOURNAL   Appl Environ Microbiol. 2002 Dec;68(12):5956-64.
  PUBMED    12450816
REFERENCE   2  (bases 1 to 7505)
  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: "Appl 
            Environ Microbiol."; date: "2002-12"; volume: "68(12)"; pages: 
            "5956-64"
COMMENT     Sequence Label: pMLBAD-eGFP
FEATURES             Location/Qualifiers
     source          1..7505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(55..60,61..77,78..83)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             410..646
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     primer_bind     670..689
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(869..1747)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1774..2058
                     /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     1981..2000
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(2093..2095,2096..2098,2099..2812)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2093..2095,2096..2098,2099..2809)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2138..2159)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2399..2418)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2746..2767
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2902..2919)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3072..3158
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3250..3277
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     4046..4065
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4132..4234)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4351..4369)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      5627..6396
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             6397..7059
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 tctacggggt ctgacgctca gtggaacgaa atcgatgagc tcgcacgaac ccagttgaca
       61 taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc
      121 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
      181 gactgttttt ttgtacagtc tagcctcggg catccaagct agctaagcgc gttacgccgt
      241 gggtcgatgt ttgatgttat ggaacagcaa cgatgttacg cagcagggta gtcgccctaa
      301 aacaaagtta ggcagccgtt gtgctggtgc tttctagtag ttgttgtggg gtaggcagtc
      361 agagctcgat ttgcttgtcg ccataataga ttcacaagaa ggattcgaca tgggtcaaag
      421 tagcgatgaa gccaacgctc ccgttgcagg gcagtttgcg cttcccctga gtgccacctt
      481 tggcttaggg gatcgcgtac gcaagaaatc tggtgccgct tggcagggtc aagtcgtcgg
      541 ttggtattgc acaaaactca ctcctgaagg ctatgcggtc gagtccgaat cccacccagg
      601 ctcagtgcaa atttatcctg tggctgcact tgaacgtgtg gcctaagcga tatcttagga
      661 tctcccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
      721 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
      781 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
      841 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
      901 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
      961 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     1021 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     1081 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     1141 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     1201 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     1261 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     1321 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     1381 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     1441 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     1501 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     1561 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     1621 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     1681 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     1741 cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca
     1801 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc
     1861 ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa
     1921 gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct
     1981 atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac
     2041 tctctactgt ttctccatac ccgttttttt gggctagcag gaggaattca ccatggtgag
     2101 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2161 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2221 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2281 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2341 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2401 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2461 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2521 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2581 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2641 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2701 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2761 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     2821 cgaattcgat cggggatcct ctagagtcga cctgcaggca tgcaagcttg gctgttttgg
     2881 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     2941 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     3001 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     3061 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     3121 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     3181 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     3241 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttccac
     3301 tcactacagc agagccattt aaacaacatc ccctccccct ttccaccgcg tcagacgccc
     3361 gtagcagccc gctacgggct ttttcatgcc ctgccctagc gtccaagcct cacggccgcg
     3421 ctcggcctct ctggcggcct tctggcgctg aggtctgcct cgtgaagaag gtgttgctga
     3481 ctcataccag gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat
     3541 gagagctttg ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg
     3601 gtctgcgttg tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat
     3661 tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta
     3721 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     3781 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     3841 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     3901 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagcga gaaatcacca
     3961 tgagtgacga ctgaatccgg tgagaatggc aaaagctaaa aaggccgtaa tatccagctg
     4021 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     4081 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     4141 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     4201 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc
     4261 ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc
     4321 cgtcacaggt atttattcga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4381 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4441 cccgcgttcc tgctggcgct gggcctgttt ctggcgctgg acttcccgct gttccgtcag
     4501 cagcttttcg cccacggcct tgatgatcgc ggcggccttg gcctgcatat cccgattcaa
     4561 cggccccagg gcgtccagaa cgggcttcag gcgctcccga aggtctcggg ccgtctcttg
     4621 ggcttgatcg gccttcttgc gcatctcacg cgctcctgcg gcggcctgta gggcaggctc
     4681 atacccctgc cgaaccgctt ttgtcagccg gtcggccacg gcttccggcg tctcaacgcg
     4741 ctttgagatt cccagctttt cggccaatcc ctgcggtgca taggcgcgtg gctcgaccgc
     4801 ttgcgggctg atggtgacgt ggcccactgg tggccgctcc agggcctcgt agaacgcctg
     4861 aatgcgcgtg tgacgtgcct tgctgccctc gatgccccgt tgcagcccta gatcggccac
     4921 agcggccgca aacgtggtct ggtcgcgggt catctgcgct ttgttgccga tgaactcctt
     4981 ggccgacagc ctgccgtcct gcgtcagcgg caccacgaac gcggtcatgt gcgggctggt
     5041 ttcgtcacgg tggatgctgg ccgtcacgat gcgatccgcc ccgtacttgt ccgccagcca
     5101 cttgtgcgcc ttctcgaaga acgccgcctg ctgttcttgg ctggccgact tccaccattc
     5161 cgggctggcc gtcatgacgt actcgaccgc caacacagcg tccttgcgcc gcttctctgg
     5221 cagcaactcg cgcagtcggc ccatcgcttc atcggtgctg ctggccgccc agtgctcgtt
     5281 ctctggcgtc ctgctggcgt cagcgttggg cgtctcgcgc tcgcggtagg cgtgcttgag
     5341 actggccgcc acgttgccca ttttcgccag cttcttgcat cgcatgatcg cgtatgccgc
     5401 catgcctgcc cctccctttt ggtgtccaac cggctcgacg ggggcagcgc aaggcggtgc
     5461 ctccggcggg ccactcaatg cttgagtata ctcactagac tttgcttcgc aaagtcgtga
     5521 ccgcctacgg cggctgcggc gccctacggg cttgctctcc gggcttcgcc ctgcgcggtc
     5581 gctgcgctcc cttgccagcc cgtggatatg tggacgatgg ccgcgagcgg ccaccggctg
     5641 gctcgcttcg ctcggcccgt ggacaaccct gctggacaag ctgatggaca ggctgcgcct
     5701 gcccacgagc ttgaccacag ggattgccca ccggctaccc agccttcgac cacataccca
     5761 ccggctccaa ctgcgcggcc tgcggccttg ccccatcaat ttttttaatt ttctctgggg
     5821 aaaagcctcc ggcctgcggc ctgcgcgctt cgcttgccgg ttggacacca agtggaaggc
     5881 gggtcaaggc tcgcgcagcg accgcgcagc ggcttggcct tgacgcgcct ggaacgaccc
     5941 aagcctatgc gagtgggggc agtcgaaggc gaagcccgcc cgcctgcccc ccgagcctca
     6001 cggcggcgag tgcgggggtt ccaagggggc agcgccacct tgggcaaggc cgaaggccgc
     6061 gcagtcgatc aacaagcccc ggaggggcca ctttttgccg gagggggagc cgcgccgaag
     6121 gcgtggggga accccgcagg ggtgcccttc tttgggcacc aaagaactag atatagggcg
     6181 aaatgcgaaa gacttaaaaa tcaacaactt aaaaaagggg ggtacgcaac agctcattgc
     6241 ggcacccccc gcaatagctc attgcgtagg ttaaagaaaa tctgtaattg actgccactt
     6301 ttacgcaacg cataattgtt gtcgcgctgc cgaaaagttg cagctgattg cgcatggtgc
     6361 cgcaaccgtg cggcacccta ccgcatggag ataagcatgg ccacgcagtc cagagaaatc
     6421 ggcattcaag ccaagaacaa gcccggtcac tgggtgcaaa cggaacgcaa agcgcatgag
     6481 gcgtgggccg ggcttattgc gaggaaaccc acggcggcaa tgctgctgca tcacctcgtg
     6541 gcgcagatgg gccaccagaa cgccgtggtg gtcagccaga agacactttc caagctcatc
     6601 ggacgttctt tgcggacggt ccaatacgca gtcaaggact tggtggccga gcgctggatc
     6661 tccgtcgtga agctcaacgg ccccggcacc gtgtcggcct acgtggtcaa tgaccgcgtg
     6721 gcgtggggcc agccccgcga ccagttgcgc ctgtcggtgt tcagtgccgc cgtggtggtt
     6781 gatcacgacg accaggacga atcgctgttg gggcatggcg acctgcgccg catcccgacc
     6841 ctgtatccgg gcgagcagca actaccgacc ggccccggcg aggagccgcc cagccagccc
     6901 ggcattccgg gcatggaacc agacctgcca gccttgaccg aaacggagga atgggaacgg
     6961 cgcgggcagc agcgcctgcc gatgcccgat gagccgtgtt ttctggacga tggcgagccg
     7021 ttggagccgc cgacacgggt cacgctgccg cgccggtagc acttgggttg cgcagcaacc
     7081 cgtaagtgcg ctgttccaga ctatcggctg tagccgcctc gccgccctat accttgtctg
     7141 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     7201 cggcacctcg ctaacggatt caccgttttt atcaggctct gggaggcaga ataaatgatc
     7261 atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca ggcatttgag
     7321 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     7381 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
     7441 tgccattcat ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagtcgacca
     7501 ataac
//
