LOCUS       sequence-53534-e        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  AUTHORS   Tett AJ, Rudder SJ, Bourdes A, Karunakaran R, Poole PS
  TITLE     Regulatable vectors for environmental gene expression in 
            alphaproteobacteria.
  JOURNAL   Appl Environ Microbiol. 2012 Oct;78(19):7137-40. Epub 2012 Jul 20.
  PUBMED    22820336
REFERENCE   2  (bases 1 to 6879)
  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: "2012-10"; volume: "78(19)"; pages: 
            "7137-40. Epub 2012 Jul 20"
COMMENT     Sequence Label: pLMB509
FEATURES             Location/Qualifiers
     source          1..6879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..884
                     /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
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             171..884
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(201..229)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     824..844
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             891..908
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      942..1028
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1717..2379)
                     /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      2380..3149
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             complement(4432..4965)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5154..5159,5160..5176,5177..5182))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tgttcacctc gcccaactgg actcatctaa cttcctatct ggatataagg caatccgaag
       61 cgtgggccta tttctattca gcaagcaaag accgtcattc aatgtacggc gcgaccccgc
      121 aaaaagcggg gcgacataac caaagagggt aacgacagga ggaagaacat atgcgtaaag
      181 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
      241 ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc
      301 ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt
      361 tcggttatgg tgttcaatgc tttgcgagat acccagatca tatgaaacgg catgactttt
      421 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
      481 ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
      541 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca
      601 actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagtta
      661 acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac
      721 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
      781 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
      841 taacagctgc tgggattaca catggcatgg atgaactata caaacatatg catcatcacc
      901 atcatcacta agagctctac aaataatgaa ttcccaggca tcaaataaaa cgaaaggctc
      961 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     1021 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     1081 caggacgccc gccataaact gccagggaat tccaactgag cgccggtcgc taccattacc
     1141 aacttgtctg gtgtcaaaaa taataggatt ttgttaaaat tcgcgttaaa tttttgttaa
     1201 atcagctcat tttttaacca ataggccgac tgcgatgagt ggcagggcgg ggcgtaattt
     1261 ttttaaggca gttattggtg cccttaaacg cctggtgcta cgcctgaata agtgataata
     1321 agcggatgaa tggcagaaat tcgaaagcaa attcgacccg gtcgtcggtt cagggcaggg
     1381 tcgttaaata gccgcttatg tctattgctg gtttaccggt ttattgacta ccggaagcag
     1441 tgtgaccgtg tgcttctcaa atgcctgagg ccagtttgct caggctctcc ccgtggaggt
     1501 aataattgac gatatgatca tttattctgc ctcccagagc ctgataaaaa cggtgaatcc
     1561 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     1621 cgcaacgcgg ggaggcagac aaggtatagg gcggcgaggc ggctacagcc gatagtctgg
     1681 aacagcgcac ttacgggttg ctgcgcaacc caagtgctac cggcgcggca gcgtgacccg
     1741 tgtcggcggc tccaacggct cgccatcgtc cagaaaacac ggctcatcgg gcatcggcag
     1801 gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc aaggctggca ggtctggttc
     1861 catgcccgga atgccgggct ggctgggcgg ctcctcgccg gggccggtcg gtagttgctg
     1921 ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca tgccccaaca gcgattcgtc
     1981 ctggtcgtcg tgatcaacca ccacggcggc actgaacacc gacaggcgca actggtcgcg
     2041 gggctggccc cacgccacgc ggtcattgac cacgtaggcc gacacggtgc cggggccgtt
     2101 gagcttcacg acggagatcc agcgctcggc caccaagtcc ttgactgcgt attggaccgt
     2161 ccgcaaagaa cgtccgatga gcttggaaag tgtcttctgg ctgaccacca cggcgttctg
     2221 gtggcccatc tgcgccacga ggtgatgcag cagcattgcc gccgtgggtt tcctcgcaat
     2281 aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc acccagtgac cgggcttgtt
     2341 cttggcttga atgccgattt ctctggactg cgtggccatg cttatctcca tgcggtaggg
     2401 tgccgcacgg ttgcggcacc atgcgcaatc agctgcaact tttcggcagc gcgacaacaa
     2461 ttatgcgttg cgtaaaagtg gcagtcaatt acagattttc tttaacctac gcaatgagct
     2521 attgcggggg gtgccgcaat gagctgttgc gtacccccct tttttaagtt gttgattttt
     2581 aagtctttcg catttcgccc tatatctagt tctttggtgc ccaaagaagg gcacccctgc
     2641 ggggttcccc cacgccttcg gcgcggctcc ccctccggca aaaagtggcc cctccggggc
     2701 ttgttgatcg actgcgcggc cttcggcctt gcccaaggtg gcgctgcccc cttggaaccc
     2761 ccgcactcgc cgccgtgagg ctcggggggc aggcgggcgg gcttcgcctt cgactgcccc
     2821 cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc aagccgctgc gcggtcgctg
     2881 cgcgagcctt gacccgcctt ccacttggtg tccaaccggc aagcgaagcg cgcaggccgc
     2941 aggccggagg cttttcccca gagaaaatta aaaaaattga tggggcaagg ccgcaggccg
     3001 cgcagttgga gccggtgggt atgtggtcga aggctgggta gccggtgggc aatccctgtg
     3061 gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt ccagcagggt tgtccacggg
     3121 ccgagcgaag cgagccagcc ggtggccgct cgcggccatc gtccacatat ccacgggctg
     3181 gcaagggagc gcagcgaccg cgcagggcga agcccggaga gcaagcccgt agggcgccgc
     3241 agccgccgta ggcggtcacg actttgcgaa gcaaagtcta gtgagtatac tcaagcattg
     3301 agtggcccgc cggaggcacc gccttgcgct gcccccgtcg agccggttgg acaccaaaag
     3361 ggaggggcag gcatggcggc atacgcgatc atgcgatgca agaagctggc gaaaatgggc
     3421 aacgtggcgg ccagtctcaa gcacgcctac cgcgagcgcg agacgcccaa cgctgacgcc
     3481 agcaggacgc cagagaacga gcactgggcg gccagcagca ccgatgaagc gatgggccga
     3541 ctgcgcgagt tgctgccaga gaagcggcgc aaggacgctg tgttggcggt cgagtacgtc
     3601 atgacggcca gcccggaatg gtggaagtcg gccagccaag aacagcaggc ggcgttcttc
     3661 gagaaggcgc acaagtggct ggcggacaag tacggggcgg atcgcatcgt gacggccagc
     3721 atccaccgtg acgaaaccag cccgcacatg accgcgttcg tggtgccgct gacgcaggac
     3781 ggcaggctgt cggccaagga gttcatcggc aacaaagcgc agatgacccg cgaccagacc
     3841 acgtttgcgg ccgctgtggc cgatctaggg ctgcaacggg gcatcgaggg cagcaaggca
     3901 cgtcacacgc gcattcaggc gttctacgag gccctggagc ggccaccagt gggccacgtc
     3961 accatcagcc cgcaagcggt cgagccacgc gcctatgcac cgcagggatt ggccgaaaag
     4021 ctgggaatct caaagcgcgt tgagacgccg gaagccgtgg ccgaccggct gacaaaagcg
     4081 gttcggcagg ggtatgagcc tgccctacag gccgccgcag gagcgcgtga gatgcgcaag
     4141 aaggccgatc aagcccaaga gacggcccga gaccttcggg agcgcctgaa gcccgttctg
     4201 gacgccctgg ggccgttgaa tcgggatatg caggccaagg ccgccgcgat catcaaggcc
     4261 gtgggcgaaa agctgctgac ggaacagcgg gaagtccagc gccagaaaca ggcccagcgc
     4321 cagcaggaac gcgggcgcgc acatttcccc gaaaagtgcc acctggcggc gttgtgacaa
     4381 tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc
     4441 ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag
     4501 agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc
     4561 gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc
     4621 gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag
     4681 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
     4741 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
     4801 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
     4861 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
     4921 agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata
     4981 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     5041 gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     5101 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac
     5161 gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc
     5221 catgggcaaa tgggattggc tttactaagc tgatccggtg gatgaccttt tgaatgacct
     5281 ttaatagatt atattactaa ttaattgggg accctagagg tccccttttt tattttaaaa
     5341 attttttcac aaaacggttt acaagcataa agcttgctca atcaatcacc ggatctagat
     5401 atcgataccc cgctccgcga tccgcgccgc agtcagggcc acgccgtcgg ctatgcggtt
     5461 gcgcgcgatc gacgagtagc ccatgcggaa gaagcggcat gggccgtcgt ccttcgggaa
     5521 gaagggcgac ccggattcga tcagcacgcc gtcctttcgc aaagcgttca tcagcaagtc
     5581 cgcatcgaga ccttccggcc cctcgatcca gaaggacgtg ccgccgaaag ccgcagatcc
     5641 ggcgatctcc aatccggcat cccggagggc agcgtccatg atgatgtggc ggcgatgata
     5701 ctcctcgcgc atgcggtgca gcacggcatc atagtgtccg agcgccagga aataggcggc
     5761 cgttctctgg aggtgccccg gcggatgccg caacatcaag gcccgtagtg cccgcgcctc
     5821 ccggatcacg gcagccggcg ccaccagata gccgagacgc aggccgggaa acaacgactt
     5881 ggagaaactg ccgatgtaga agacccggcc gcagcggtcg aaggccttga gggccgggga
     5941 ggggggcgcc agaaagctca tctcgaactc gtaatcgtcc tcgacgatta tgaagtcctt
     6001 ggaggcggcc gcctcgagca aggccgcccg ccgttcgatc ggcatcgtcg cgccggtcgg
     6061 cgaatgatgg ctgggcgtga cgaacacggc atcgacctct tctggaagcg cggcgggcgg
     6121 caggccgtcc ttgtccacgt cgacgaaggt gatcccggcg ccgctcaagc gaagcgatgc
     6181 actcatgtcg ggatggcagg ggttctcgca gaccgcatgg gcgccctgac gcaacaggag
     6241 ctggatgacg atccagaggg cgttctgcgc gcccaccgtc accaggatct cgtcggggtt
     6301 cgcgcggatg ccgcggcccg gcaatgtgcg cgagcagata tagttgacaa gctggatgtc
     6361 gtccgccgcc gcgaagtcgc ccgccatcag ctcgaaatcc tcgcgcgcca gcgcccgtcg
     6421 cgcgcagtcg cgccaggccg tgaggtcgaa cagcgacggg tccatctgac cgtataggaa
     6481 cggataaggg taggttcgcc agtcgagggg cttacgcatc tgcttggcca cgatgaaggt
     6541 cgagcgcaac ttggccgacc agtcgatggc atcggccgca gcatgggcgg cctgcatcgc
     6601 gatggctgtc gttgggggct tgccggccac acggtagccg ctccggttcg ccgcctccac
     6661 atagccctgg gaagcgagtt cctgataggc gagcgttacg gtgatccgcg aaatattcag
     6721 ataggccgca agcttgcggc tggagggcaa ttgggcgccc ggcgccaggc ggcccgacaa
     6781 tacggccgaa acgatcgttt cgcggatctg cgcctgcagg cccgtttggc tggcccggtc
     6841 aaggaaaaag atcgtttcgg agacggtcga gcccgccat
//
