LOCUS       sequence-224237-        6922 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 6922)
  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 6922)
  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: pMLS7-egfp
FEATURES             Location/Qualifiers
     source          1..6922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..720)
                     /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(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(809..826)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      979..1065
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1157..1184
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1650..2312)
                     /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      2313..3084
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     primer_bind     4342..4360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4477..4579
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4646..4665)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(5434..5670)
                     /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"
     promoter        complement(join(5996..6001,6002..6018,6019..6024))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
      721 agcggccgcg aattcgatcg ggatcctcta gagtcgacct gcaggcatgc aagcttggct
      781 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
      841 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
      901 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
      961 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1021 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     1081 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     1141 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     1201 cttgttattg gtcgacttaa acgcctggtg ctacgcctga ataagtgata ataagcggat
     1261 gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa
     1321 atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc
     1381 gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt
     1441 gacgatatga tcatttattc tgcctcccag agcctgataa aaacggtgaa tccgttagcg
     1501 aggtgccgcc ggcttccatt caggtcgagg cggcccggct ccatgcaccg cgacgcaacg
     1561 cggggaggca gacaaggtat agggcggcga ggcggctaca gccgatagtc tggaacagcg
     1621 cacttacggg ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc
     1681 ggctccaacg gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc
     1741 tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc
     1801 ggaatgccgg gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc
     1861 ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg
     1921 tcgtgatcaa ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg
     1981 ccccacgcca cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc
     2041 acgacggaga tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa
     2101 gaacgtccga tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc
     2161 atctgcgcca cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg
     2221 gcccacgcct catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct
     2281 tgaatgccga tttctctgga ctgcgtggcc atgcttatct ccatgcggta ggggtgccgc
     2341 acggttgcgg caccatgcgc aatcagctgc aacttttcgg cagcgcgaca acaattatgc
     2401 gttgcgtaaa agtggcagtc aattacagat tttctttaac ctacgcaatg agctattgcg
     2461 gggggtgccg caatgagctg ttgcgtaccc ccctttttta agttgttgat ttttaagtct
     2521 ttcgcatttc gccctatatc tagttctttg gtgcccaaag aagggcaccc ctgcggggtt
     2581 cccccacgcc ttcggcgcgg ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg
     2641 atcgactgcg cggccttcgg ccttgcccaa ggtggcgctg cccccttgga acccccgcac
     2701 tcgccgccgt gaggctcggg gggcaggcgg gcgggcttcg cccttcgact gcccccactc
     2761 gcataggctt gggtcgttcc aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga
     2821 gccttgaccc gccttccact tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc
     2881 ggaggctttt ccccagagaa aattaaaaaa attgatgggg caaggccgca ggccgcgcag
     2941 ttggagccgg tgggtatgtg gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa
     3001 gctcgtgggc aggcgcagcc tgtccatcag cttgtccagc agggttgtcc acgggccgag
     3061 cgaagcgagc cagccggtgg ccgctcgcgg ccatcgtcca catatccacg ggctggcaag
     3121 ggagcgcagc gaccgcgcag ggcgaagccc ggagagcaag cccgtagggc gccgcagccg
     3181 ccgtaggcgg tcacgacttt gcgaagcaaa gtctagtgag tatactcaag cattgagtgg
     3241 cccgccggag gcaccgcctt gcgctgcccc cgtcgagccg gttggacacc aaaagggagg
     3301 ggcaggcatg gcggcatacg cgatcatgcg atgcaagaag ctggcgaaaa tgggcaacgt
     3361 ggcggccagt ctcaagcacg cctaccgcga gcgcgagacg cccaacgctg acgccagcag
     3421 gacgccagag aacgagcact gggcggccag cagcaccgat gaagcgatgg gccgactgcg
     3481 cgagttgctg ccagagaagc ggcgcaagga cgctgtgttg gcggtcgagt acgtcatgac
     3541 ggccagcccg gaatggtgga agtcggccag ccaagaacag caggcggcgt tcttcgagaa
     3601 ggcgcacaag tggctggcgg acaagtacgg ggcggatcgc atcgtgacgg ccagcatcca
     3661 ccgtgacgaa accagcccgc acatgaccgc gttcgtggtg ccgctgacgc aggacggcag
     3721 gctgtcggcc aaggagttca tcggcaacaa agcgcagatg acccgcgacc agaccacgtt
     3781 tgcggccgct gtggccgatc tagggctgca acggggcatc gagggcagca aggcacgtca
     3841 cacgcgcatt caggcgttct acgaggccct ggagcggcca ccagtgggcc acgtcaccat
     3901 cagcccgcaa gcggtcgagc cacgcgccta tgcaccgcag ggattggccg aaaagctggg
     3961 aatctcaaag cgcgttgaga cgccggaagc cgtggccgac cggctgacaa aagcggttcg
     4021 gcaggggtat gagcctgccc tacaggccgc cgcaggagcg cgtgagatgc gcaagaaggc
     4081 cgatcaagcc caagagacgg cccgagacct tcgggagcgc ctgaagcccg ttctggacgc
     4141 cctggggccg ttgaatcggg atatgcaggc caaggccgcc gcgatcatca aggccgtggg
     4201 cgaaaagctg ctgacggaac agcgggaagt ccagcgccag aaacaggccc agcgccagca
     4261 ggaacgcggg cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4321 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcgaataaat
     4381 acctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg
     4441 gaagccctgg gccaactttt ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa
     4501 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     4561 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     4621 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     4681 ccagaccgtt cagctggata ttacggcctt tttagctttt gccattctca ccggattcag
     4741 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     4801 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     4861 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     4921 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     4981 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     5041 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga
     5101 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa
     5161 caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc
     5221 ctggtatgag tcagcaacac cttcttcacg aggcagacct cagcgccaga aggccgccag
     5281 agaggccgag cgcggccgtg aggcttggac gctagggcag ggcatgaaaa agcccgtagc
     5341 gggctgctac gggcgtctga cgcggtggaa agggggaggg gatgttgttt aaatggctct
     5401 gctgtagtga gtggagggat cctaagatat cgcttaggcc acacgttcaa gtgcagccac
     5461 aggataaatt tgcactgagc ctgggtggga ttcggactcg accgcatagc cttcaggagt
     5521 gagttttgtg caataccaac cgacgacttg accctgccaa gcggcaccag atttcttgcg
     5581 tacgcgatcc cctaagccaa aggtggcact caggggaagc gcaaactgcc ctgcaacggg
     5641 agcgttggct tcatcgctac tttgacccat gtcgaatcct tcttgtgaat ctattatggc
     5701 gacaagcaaa tcgagctctg actgcctacc ccacaacaac tatcagaaag caccagcaca
     5761 acggctgcct aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca
     5821 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgtcc gaggcattga
     5881 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
     5941 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
     6001 acgaaccgaa caggcttatg tcaactgggt tcgtgcgagc tcatcgattt cgttccactg
     6061 agcgtcagac cccgtagata cagctagccg aattgctgga agtcgtcgcc gccgactaag
     6121 cgctcagcga gcgcctggcc aggtcggcag gggtacgaaa catcgcaacc tgttgactcc
     6181 cttagtaatt tcggaatatt atgctaggtt ccggttcttg gaatgcctgc gcgcgaagtc
     6241 tattcgtgcg ccgactaccg gtccggaagt tcgatgcgca atcgtttctg ctttgcccgg
     6301 aaacagatgc gtcgattttg ttcaatttca ggaataaaca atgccaacca tcaatcaact
     6361 ggttcgcaaa ggccgcgcgt cggaaacgac gaagagcaag agcccggctt tgcaggactg
     6421 cccccagcgt cgcggcgtgt gcacccgtgt gtacaccacg acgccgaaga agcctaactc
     6481 ggcactgcgt aaggttgcca aggttcgtct gacgaacggc ttcgaagtga tttcttacat
     6541 cggtggtgaa ggccacaatc tgcaggaaca ctcggtcgtg ctgattcgcg gcggtcgtgt
     6601 caaggacttg ccgggtgtgc gttaccacat ggttcgtggc tcgctggata cccagggcgt
     6661 caaggatcgt aagcaggctc gctcgaagta cggtgcgaag cgtgcaaaag ctggtaaata
     6721 agctggcaac caattagccg gtcagtagtt tcaggtcgcc tgcaaaggcg gtaatagcgg
     6781 tggtgccgga atcgccggtg ctgtcgagta agtggtcacc cgaccaggct ggtaagtcgt
     6841 tagggatgaa tcgggttagt tggtggccgc ggggctaaaa gttagctcca actgaaaagt
     6901 taaaggaaga aacgaattca cc
//
