LOCUS       sequence-376365-        7460 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 7460)
  AUTHORS   Miller WG, Leveau JH, Lindow SE
  TITLE     Improved gfp and inaZ broad-host-range promoter-probe vectors.
  JOURNAL   Mol Plant Microbe Interact. 2000 Nov;13(11):1243-50.
  PUBMED    11059491
REFERENCE   2  (bases 1 to 7460)
  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: "Mol Plant 
            Microbe Interact."; date: "2000-11"; volume: "13(11)"; pages: 
            "1243-50"
COMMENT     Sequence Label: pPROBE-OT
FEATURES             Location/Qualifiers
     source          1..7460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     473..495
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     487..504
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     488..504
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     514..533
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        514..532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      575..661
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      758..844
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      941..1027
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1124..1210
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1302..1358
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             1409..2122
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1439..1467)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2062..2082
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2143..2229
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3186..3977
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5805..6576
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             6577..7239
                     /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 cacggggaga gcctgagcaa actggcctca ggcatttgag aagcacacgg tcacactgct
       61 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
      121 ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta
      181 tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc ttaaaaaaat
      241 tacgccccgc cctgccactc atcgcagtac ggcctattgg ttaaaaaatg agctgattta
      301 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
      361 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      421 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      481 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
      541 gagctggggc cgcaattccc aattccccag gcatcaaata aaacgaaagg ctcagtcgaa
      601 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
      661 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
      721 cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa aggctcagtc
      781 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      841 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
      901 acgcccgcca taaactgcca ggaattaatt ccccaggcat caaataaaac gaaaggctca
      961 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1021 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     1081 aggacgcccg ccataaactg ccaggaatta attccccagg catcaaataa aacgaaaggc
     1141 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1201 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     1261 ggcaggacgc ccgccataaa ctgccaggaa ttggggatcg gaagcttgca tgcctgcagg
     1321 tcgactctag aggatccccg ggtaccgagc tcgaattcgg cttattccct aactaactaa
     1381 agattaactt tataaggagg aaaaacatat gagtaaagga gaagaacttt tcactggagt
     1441 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     1501 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     1561 aaaactacct gttccatggc caacacttgt cactactttg acttatggtg ttcaatgctt
     1621 ttcaagatac ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg
     1681 ttatgtacag gaaagaacta tatttttcaa agatgacggg aactataaga cacgtgctga
     1741 agtcaagttt gaaggtgata cacttgttaa tagaatcgag ttaaaaggta ttgattttaa
     1801 agaagatgga aacattcttg gacacaagtt ggaatacaac tataactcac acaatgtata
     1861 catcatggca gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat
     1921 tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     1981 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
     2041 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     2101 tggcatggat gaactataca aataggaatt aattccaggc atcaaataaa acgaaaggct
     2161 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     2221 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     2281 gcaggacgcc cgccataaac tgccaggaat tgggggcggc cccccctcga ggtcagcttt
     2341 atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa aaaggggacc tctagggtcc
     2401 ccaattaatt agtaatataa tctattaaag gtcattcaaa aggtcatcca ccggatcaat
     2461 tcccctgctc gcgcaggctg ggtgccaagc tctcgggtaa catcaaggcc cgatccttgg
     2521 agcccttgcc ctcccgcacg atgatcgtgc cgtgatcgaa atccagatcc ttgacccgca
     2581 gttgcaaacc ctcactgatc cgcatgcccg ttccatacag aagctgggcg aacaaacgat
     2641 gctcgccttc cagaaaaccg aggatgcgaa ccacttcatc cggggtcagc accaccggca
     2701 agcgccgcga cggccgaggt cttccgatct cctgaagcca gggcagatcc gtgcacagca
     2761 ccttgccgta gaagaacagc aaggccgcca atgcctgacg atgcgtggag accgaaacct
     2821 tgcgctcgtt cgccagccag gacagaaatg cctcgacttc gctgctgccc aaggttgccg
     2881 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
     2941 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     3001 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     3061 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     3121 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     3181 acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     3241 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     3301 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     3361 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     3421 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     3481 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     3541 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     3601 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     3661 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     3721 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     3781 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     3841 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     3901 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     3961 aggtagtcgg caaataatgt ctaacaattc gttcaagccg acgccgcttc gcggcgcggc
     4021 ttaactcaag cgttagatgc actaagcaca taattgctca cagccaaact atcaggtcaa
     4081 gtctgctttt attattttta agcgtgcata ataagcccta cacaaattgg gagatatatc
     4141 atgaaaggct ggctttttct tgttatcgca atagttggcg aagtaatcgc aacatccgca
     4201 ttaaaatcta gcgagggctt tactaagctg atccggtgga tgaccttttg aatgaccttt
     4261 aatagattat attactaatt aattggggac cctagaggtc ccctttttta ttttaaaaat
     4321 tttttcacaa aacggtttac aagcataaag ctgggggatc ttgatcccct gcgccatcag
     4381 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     4441 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     4501 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     4561 cagatagccc agtagctgac attcatccca ggtggcactt ttcggggaaa tgtgcgcgcc
     4621 cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt tccgtcagca
     4681 gcttttcgcc cacggccttg atgatcgcgg cggccttggc ctgcatatcc cgattcaacg
     4741 gccccagggc gtccagaacg ggcttcaggc gctcccgaag gtctcgggcc gtctcttggg
     4801 cttgatcggc cttcttgcgc atctcacgcg ctcctgcggc ggcctgtagg gcaggctcat
     4861 acccctgccg aaccgctttt gtcagccggt cggccacggc ttccggcgtc tcaacgcgct
     4921 ttgagattcc cagcttttcg gccaatccct gcggtgcata ggcgcgtggc tcgaccgctt
     4981 gcgggctgat ggtgacgtgg cccactggtg gccgctccag ggcctcgtag aacgcctgaa
     5041 tgcgcgtgtg acgtgccttg ctgccctcga tgccccgttg cagccctaga tcggccacag
     5101 cggccgcaaa cgtggtctgg tcgcgggtca tctgcgcttt gttgccgatg aactccttgg
     5161 ccgacagcct gccgtcctgc gtcagcggca ccacgaacgc ggtcatgtgc gggctggttt
     5221 cgtcacggtg gatgctggcc gtcacgatgc gatccgcccc gtacttgtcc gccagccact
     5281 tgtgcgcctt ctcgaagaac gccgcctgct gttcttggct ggccgacttc caccattccg
     5341 ggctggccgt catgacgtac tcgaccgcca acacagcgtc cttgcgccgc ttctctggca
     5401 gcaactcgcg cagtcggccc atcgcttcat cggtgctgct ggccgcccag tgctcgttct
     5461 ctggcgtcct gctggcgtca gcgttgggcg tctcgcgctc gcggtaggcg tgcttgagac
     5521 tggccgccac gttgcccatt ttcgccagct tcttgcatcg catgatcgcg tatgccgcca
     5581 tgcctgcccc tcccttttgg tgtccaaccg gctcgacggg ggcagcgcaa ggcggtgcct
     5641 ccggcgggcc actcaatgct tgagtatact cactagactt tgcttcgcaa agtcgtgacc
     5701 gcctacggcg gctgcggcgc cctacgggct tgctctccgg gcttcgccct gcgcggtcgc
     5761 tgcgctccct tgccagcccg tggatatgtg gacgatggcc gcgagcggcc accggctggc
     5821 tcgcttcgct cggcccgtgg acaaccctgc tggacaagct gatggacagg ctgcgcctgc
     5881 ccacgagctt gaccacaggg attgcccacc ggctacccag ccttcgacca catacccacc
     5941 ggctccaact gcgcggcctg cggccttgcc ccatcaattt ttttaatttt ctctggggaa
     6001 aagcctccgg cctgcggcct gcgcgcttcg cttgccggtt ggacaccaag tggaaggcgg
     6061 gtcaaggctc gcgcagcgac cgcgcagcgg cttggccttg acgcgcctgg aacgacccaa
     6121 gcctatgcga gtgggggcag tcgaagggcg aagcccgccc gcctgccccc cgagcctcac
     6181 ggcggcgagt gcgggggttc caagggggca gcgccacctt gggcaaggcc gaaggccgcg
     6241 cagtcgatca acaagccccg gaggggccac tttttgccgg agggggagcc gcgccgaagg
     6301 cgtgggggaa ccccgcaggg gtgcccttct ttgggcacca aagaactaga tatagggcga
     6361 aatgcgaaag acttaaaaat caacaactta aaaaaggggg gtacgcaaca gctcattgcg
     6421 gcaccccccg caatagctca ttgcgtaggt taaagaaaat ctgtaattga ctgccacttt
     6481 tacgcaacgc ataattgttg tcgcgctgcc gaaaagttgc agctgattgc gcatggtgcc
     6541 gcaaccgtgc ggcaccccta ccgcatggag ataagcatgg ccacgcagtc cagagaaatc
     6601 ggcattcaag ccaagaacaa gcccggtcac tgggtgcaaa cggaacgcaa agcgcatgag
     6661 gcgtgggccg ggcttattgc gaggaaaccc acggcggcaa tgctgctgca tcacctcgtg
     6721 gcgcagatgg gccaccagaa cgccgtggtg gtcagccaga agacactttc caagctcatc
     6781 ggacgttctt tgcggacggt ccaatacgca gtcaaggact tggtggccga gcgctggatc
     6841 tccgtcgtga agctcaacgg ccccggcacc gtgtcggcct acgtggtcaa tgaccgcgtg
     6901 gcgtggggcc agccccgcga ccagttgcgc ctgtcggtgt tcagtgccgc cgtggtggtt
     6961 gatcacgacg accaggacga atcgctgttg gggcatggcg acctgcgccg catcccgacc
     7021 ctgtatccgg gcgagcagca actaccgacc ggccccggcg aggagccgcc cagccagccc
     7081 ggcattccgg gcatggaacc agacctgcca gccttgaccg aaacggagga atgggaacgg
     7141 cgcgggcagc agcgcctgcc gatgcccgat gagccgtgtt ttctggacga tggcgagccg
     7201 ttggagccgc cgacacgggt cacgctgccg cgccggtagc acttgggttg cgcagcaacc
     7261 cgtaagtgcg ctgttccaga ctatcggctg tagccgcctc gccgccctat accttgtctg
     7321 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     7381 cggcacctcg ctaacggatt caccgttttt atcaggctct gggaggcaga ataaatgatc
     7441 atatcgtcaa ttattacctc
//
