LOCUS       sequence-8098-ex        5384 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 5384)
  AUTHORS   Karunakaran R, Mauchline TH, Hosie AH, Poole PS
  TITLE     A family of promoter probe vectors incorporating autofluorescent and
            chromogenic reporter proteins for studying gene expression in 
            Gram-negative bacteria.
  JOURNAL   Microbiology. 2005 Oct . 151(Pt 10):3249-56.
  PUBMED    16207908
REFERENCE   2  (bases 1 to 5384)
  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: 
            "Microbiology. 2005 Oct . 151(Pt 10):3249-56."
COMMENT     Sequence Label: pRU1099
FEATURES             Location/Qualifiers
     source          1..5384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             326..1039
                     /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             326..1039
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(356..384)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     979..999
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1107..1193
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1882..2544)
                     /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      2545..3314
                     /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(4597..5130)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5319..5324,5325..5341,5342..5347))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ccatgggcaa atgggattgg ctttactaag ctgatccggt ggatgacctt ttgaatgacc
       61 tttaatagat tatattacta attaattggg gaccctagag gtcccctttt ttattttaaa
      121 aattttttca caaaacggtt tacaagcata aagcttgctc aatcaatcac cggatctaga
      181 tatcgattaa ttaagtcgac atctagagtt taaactttaa ttaagcccgg gctgcagccc
      241 gggggatcca ctaggttaac gcatgcgggc ccggtaccag gcctagatag atagagagag
      301 agagagacta gtggaggaag aaaaaatgcg taaaggagaa gaacttttca ctggagttgt
      361 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
      421 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
      481 actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc aatgctttgc
      541 gagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
      601 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
      661 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
      721 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
      781 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga
      841 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
      901 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
      961 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     1021 catggatgaa ctatacaaag cagcaaacga cgaaaactac gctgcagcag tttaatgagc
     1081 tctacaaata atgaattccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1141 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     1201 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     1261 aaactgccag ggaattccaa ctgagcgccg gtcgctacca ttaccaactt gtctggtgtc
     1321 aaaaataata ggattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1381 aaccaatagg ccgactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     1441 tggtgccctt aaacgcctgg tgctacgcct gaataagtga taataagcgg atgaatggca
     1501 gaaattcgaa agcaaattcg acccggtcgt cggttcaggg cagggtcgtt aaatagccgc
     1561 ttatgtctat tgctggttta ccggtttatt gactaccgga agcagtgtga ccgtgtgctt
     1621 ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg gaggtaataa ttgacgatat
     1681 gatcatttat tctgcctccc agagcctgat aaaaacggtg aatccgttag cgaggtgccg
     1741 ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     1801 cagacaaggt atagggcggc gaggcggcta cagccgatag tctggaacag cgcacttacg
     1861 ggttgctgcg caacccaagt gctaccggcg cggcagcgtg acccgtgtcg gcggctccaa
     1921 cggctcgcca tcgtccagaa aacacggctc atcgggcatc ggcaggcgct gctgcccgcg
     1981 ccgttcccat tcctccgttt cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc
     2041 gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc ccggatacag
     2101 ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc
     2161 aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct ggccccacgc
     2221 cacgcggtca ttgaccacgt aggccgacac ggtgccgggg ccgttgagct tcacgacgga
     2281 gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca aagaacgtcc
     2341 gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc ccatctgcgc
     2401 cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc cggcccacgc
     2461 ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc
     2521 gatttctctg gactgcgtgg ccatgcttat ctccatgcgg tagggtgccg cacggttgcg
     2581 gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg cgttgcgtaa
     2641 aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc ggggggtgcc
     2701 gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc tttcgcattt
     2761 cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc
     2821 cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc
     2881 gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca ctcgccgccg
     2941 tgaggctcgg ggggcaggcg ggcgggcttc gccttcgact gcccccactc gcataggctt
     3001 gggtcgttcc aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc
     3061 gccttccact tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt
     3121 ccccagagaa aattaaaaaa attgatgggg caaggccgca ggccgcgcag ttggagccgg
     3181 tgggtatgtg gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc
     3241 aggcgcagcc tgtccatcag cttgtccagc agggttgtcc acgggccgag cgaagcgagc
     3301 cagccggtgg ccgctcgcgg ccatcgtcca catatccacg ggctggcaag ggagcgcagc
     3361 gaccgcgcag ggcgaagccc ggagagcaag cccgtagggc gccgcagccg ccgtaggcgg
     3421 tcacgacttt gcgaagcaaa gtctagtgag tatactcaag cattgagtgg cccgccggag
     3481 gcaccgcctt gcgctgcccc cgtcgagccg gttggacacc aaaagggagg ggcaggcatg
     3541 gcggcatacg cgatcatgcg atgcaagaag ctggcgaaaa tgggcaacgt ggcggccagt
     3601 ctcaagcacg cctaccgcga gcgcgagacg cccaacgctg acgccagcag gacgccagag
     3661 aacgagcact gggcggccag cagcaccgat gaagcgatgg gccgactgcg cgagttgctg
     3721 ccagagaagc ggcgcaagga cgctgtgttg gcggtcgagt acgtcatgac ggccagcccg
     3781 gaatggtgga agtcggccag ccaagaacag caggcggcgt tcttcgagaa ggcgcacaag
     3841 tggctggcgg acaagtacgg ggcggatcgc atcgtgacgg ccagcatcca ccgtgacgaa
     3901 accagcccgc acatgaccgc gttcgtggtg ccgctgacgc aggacggcag gctgtcggcc
     3961 aaggagttca tcggcaacaa agcgcagatg acccgcgacc agaccacgtt tgcggccgct
     4021 gtggccgatc tagggctgca acggggcatc gagggcagca aggcacgtca cacgcgcatt
     4081 caggcgttct acgaggccct ggagcggcca ccagtgggcc acgtcaccat cagcccgcaa
     4141 gcggtcgagc cacgcgccta tgcaccgcag ggattggccg aaaagctggg aatctcaaag
     4201 cgcgttgaga cgccggaagc cgtggccgac cggctgacaa aagcggttcg gcaggggtat
     4261 gagcctgccc tacaggccgc cgcaggagcg cgtgagatgc gcaagaaggc cgatcaagcc
     4321 caagagacgg cccgagacct tcgggagcgc ctgaagcccg ttctggacgc cctggggccg
     4381 ttgaatcggg atatgcaggc caaggccgcc gcgatcatca aggccgtggg cgaaaagctg
     4441 ctgacggaac agcgggaagt ccagcgccag aaacaggccc agcgccagca ggaacgcggg
     4501 cgcgcacatt tccccgaaaa gtgccacctg gcggcgttgt gacaatttac cgaacaactc
     4561 cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat
     4621 atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc
     4681 gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt
     4741 gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag
     4801 atcatagata tagatctcac tacgcggctg ctcaaacctg ggcagaacgt aagccgcgag
     4861 agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag
     4921 caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa
     4981 ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct
     5041 acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg
     5101 ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac
     5161 ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt
     5221 cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc
     5281 tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt
     5341 atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgt
//
