LOCUS       sequence-60702-e        9323 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 9323)
  AUTHORS   Anthouard R, DiRita VJ
  TITLE     Small-molecule inhibitors of toxT expression in Vibrio cholerae.
  JOURNAL   MBio. 2013 Aug 6;4(4). pii: e00403-13. doi: 10.1128/mBio.00403-13.
  PUBMED    23919997
REFERENCE   2  (bases 1 to 9323)
  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: "MBio."; 
            date: "2013-08-6"; volume: "4(4). pii: e00403-13. doi"; pages: " 
            10.1128/mBio.00403-13"
COMMENT     Sequence Label: pMMB66EH-aphA
FEATURES             Location/Qualifiers
     source          1..9323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(111..179,180..971)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(329..348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1408..1802
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            2144..2231
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             3470..4441
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDAPTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGHDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             4952..5791
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             5778..6629
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRKRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     primer_bind     7949..7968
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7988..8065)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        join(8295..8300,8301..8316,8317..8323)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    8331..8347
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8336..8358
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(8948..8965)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      9118..9204
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      9296..9323
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
       61 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      121 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      181 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
      241 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
      301 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
      361 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
      421 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
      481 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
      541 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
      601 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
      661 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
      721 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
      781 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
      841 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
      901 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
      961 agcattggta actgtcagac caagtttact catatatact ttagattgat ttctgaaagc
     1021 gaccaggtgc tcggcgtggc aagactcgca gcgaacccgt agaaagccat gctccagccg
     1081 cccgcattgg agaaattctt caaattcccg ttgcacatag cccggcaatt cctttccctg
     1141 ctctgccata agcgcagcga atgccgggta atactcgtca acgatctgat agagaagggt
     1201 ttgctcgggt cggtggctct ggtaacgacc agtatcccga tcccggctgg ccgtcctggc
     1261 cgccacatga ggcatgttcc gcgtccttgc aatactgtgt ttacatacag tctatcgctt
     1321 agcggaaagt tcttttaccc tcagccgaaa tgcctgccgt tgctagacat tgccagccag
     1381 tgcccgtcac tcccgtacta actgtcacga acccctgcaa taactgtcac gcccccctgc
     1441 aataactgtc acgaacccct gcaataactg tcacgccccc aaacctgcaa acccagcagg
     1501 ggcgggggct ggcggggtgt tggaaaaatc catccatgat tatctaagaa taatccacta
     1561 ggcgcggtta tcagcgccct tgtggggcgc tgctgccctt gcccaatatg cccggccaga
     1621 ggccggatag ctggtctatt cgctgcgcta ggctacacac cgccccaccg ctgcgcggca
     1681 gggggaaagg cgggcaaagc ccgctaaacc ccacaccaaa ccccgcagaa atacgctgga
     1741 gcgcttttag ccgctttagc ggcctttccc cctacccgaa gggtgggggc gcgtgtgcag
     1801 ccccgcaggg cctgtctcgg tcgatcattc agcccggctc atccttctgg cgtggcggca
     1861 gaccgaacaa ggcgcggtcg tggtcgcgtt caaggtacgc atccattgcc gccatgagcc
     1921 gatcctccgg ccactcgctg ctgttcacct tggccaaaat catggccccc accagcacct
     1981 tgcgccttgt ttcgttcttg cgctcttgct gctgttccct tgcccgctcc cgctgaattt
     2041 cggcattgat tcgcgctcgt tgttcttcga gcttggccag ccgatccgcc gccttgttgc
     2101 tccccttaac catcttgaca ccccattgtt aatgtgctgt ctcgtaggct atcatggagg
     2161 cacagcggcg gcaatcccga ccctactttg taggggaggg cgcacttacc ggtttctctt
     2221 cgagaaactg gcctaacggc cacccttcgg gcggtgcgct ctccgagggc cattgcatgg
     2281 agccgaaaag caaaagcaac agcgaggcag catggcgatt tatcacctta cggcgaaaac
     2341 cggcagcagg tcgggcggcc aatcggccag ggccaaggcc gactacatcc agcgcgaagg
     2401 caagtatgcc cgcgacatgg atgaagtctt gcacgccgaa tccgggcaca tgccggagtt
     2461 cgtcgagcgg cccgccgact actgggatgc tgccgacctg tatgaacgcg ccaatgggcg
     2521 gctgttcaag gaggtcgaat ttgccctgcc ggtcgagctg accctcgacc agcagaaggc
     2581 gctggcgtcc gagttcgccc agcacctgac cggtgccgag cgcctgccgt atacgctggc
     2641 catccatgcc ggtggcggcg agaacccgca ctgccacctg atgatctccg agcggatcaa
     2701 tgacggcatc gagcggcccg ccgctcagtg gttcaagcgg tacaacggca agaccccgga
     2761 gaagggcggg gcacagaaga ccgaagcgct caagcccaag gcatggcttg agcagacccg
     2821 cgaggcatgg gccgaccatg ccaaccgggc attagagcgg gctggccacg acgcccgcat
     2881 tgaccacaga acacttgagg cgcagggcat cgagcgcctg cccggtgttc acctggggcc
     2941 gaacgtggtg gagatggaag gccggggcat ccgcaccgac cgggcagacg tggccctgaa
     3001 catcgacacc gccaacgccc agatcatcga cttacaggaa taccgggagg caatagacca
     3061 tgaacgcaat cgacagagtg aagaaatcca gaggcatcaa cgagttagcg gagcagatcg
     3121 aaccgctggc ccagagcatg gcgacactgg ccgacgaagc ccggcaggtc atgagccaga
     3181 ccaagcaggc cagcgaggcg caggcggcgg agtggctgaa agcccagcgc cagacagggg
     3241 cggcatgggt ggagctggcc aaagagttgc gggaggtagc cgccgaggtg agcagcgccg
     3301 cgcagagcgc ccggagcgcg tcgcgggggt ggcactggaa gctatggcta accgtgatgc
     3361 tggcttccat gatgcctacg gtggtgctgc tgatcgcatc gttgctcttg ctcgacctga
     3421 cgccactgac aaccgaggac ggctcgatct ggctgcgctt ggtggcccga tgaagaacga
     3481 caggactttg caggccatag gccgacagct caaggccatg ggctgtgagc gcttcgatat
     3541 cggcgtcagg gacgcaccca ccggccagat gatgaaccgg gaatggtcag ccgccgaagt
     3601 gctccagaac acgccatggc tcaagcggat gaatgcccag ggcaatgacg tgtatatcag
     3661 gcccgccgag caggagcggc atggtctggt gctggtggac gacctcagcg agtttgacct
     3721 ggatgacatg aaagccgagg gccgggagcc tgccctggta gtggaaacca gcccgaagaa
     3781 ctatcaggca tgggtcaagg tggccgacgc cgcaggcggt gaacttcggg ggcagattgc
     3841 ccggacgctg gccagcgagt acgacgccga cccggccagc gccgacagcc gccactatgg
     3901 ccgcttggcg ggcttcacca accgcaagga caagcacacc acccgcgccg gttatcagcc
     3961 gtgggtgctg ctgcgtgaat ccaagggcaa gaccgccacc gctggcccgg cgctggtgca
     4021 gcaggctggc cagcagatcg agcaggccca gcggcagcag gagaaggccc gcaggctggc
     4081 cagcctcgaa ctgcccgagc ggcagcttag ccgccaccgg cgcacggcgc tggacgagta
     4141 ccgcagcgag atggccgggc tggtcaagcg cttcggtcat gacctcagca agtgcgactt
     4201 tatcgccgcg cagaagctgg ccagccgggg ccgcagtgcc gaggaaatcg gcaaggccat
     4261 ggccgaggcc agcccagcgc tggcagagcg caagcccggc cacgaagcgg attacatcga
     4321 gcgcaccgtc agcaaggtca tgggtctgcc cagcgtccag cttgcgcggg ccgagctggc
     4381 acgggcaccg gcaccccgcc agcgaggcat ggacaggggc gggccagatt tcagcatgta
     4441 gtgcttgcgt tggtactcac gcctgttata ctatgagtac tcacgcacag aagggggttt
     4501 tatggaatac gaaaaaagcg cttcagggtc ggtctacctg atcaaaagtg acaagggcta
     4561 ttggttgccc ggtggctttg gttatacgtc aaacaaggcc gaggctggcc gcttttcagt
     4621 cgctgatatg gccagcctta accttgacgg ctgcaccttg tccttgttcc gcgaagacaa
     4681 gcctttcggc cccggcaagt ttctcggtga ctgatatgaa agaccaaaag gacaagcaga
     4741 ccggcgacct gctggccagc cctgacgctg tacgccaagc gcgatatgcc gagcgcatga
     4801 aggccaaagg gatgcgtcag cgcaagttct ggctgaccga cgacgaatac gaggcgctgc
     4861 gcgagtgcct ggaagaactc agagcggcgc agggcggggg tagtgacccc gccagcgcct
     4921 aaccaccaac tgcctgcaaa ggaggcaatc aatggctacc cataagccta tcaatattct
     4981 ggaggcgttc gcagcagcgc cgccaccgct ggactacgtt ttgcccaaca tggtggccgg
     5041 tacggtcggg gcgctggtgt cgcccggtgg tgccggtaaa tccatgctgg ccctgcaact
     5101 ggccgcacag attgcaggcg ggccggatct gctggaggtg ggcgaactgc ccaccggccc
     5161 ggtgatctac ctgcccgccg aagacccgcc caccgccatt catcaccgcc tgcacgccct
     5221 tggggcgcac ctcagcgccg aggaacggca agccgtggct gacggcctgc tgatccagcc
     5281 gctgatcggc agcctgccca acatcatggc cccggagtgg ttcgacggcc tcaagcgcgc
     5341 cgccgagggc cgccgcctga tggtgctgga cacgctgcgc cggttccaca tcgaggaaga
     5401 aaacgccagc ggccccatgg cccaggtcat cggtcgcatg gaggccatcg ccgccgatac
     5461 cgggtgctct atcgtgttcc tgcaccatgc cagcaagggc gcggccatga tgggcgcagg
     5521 cgaccagcag caggccagcc ggggcagctc ggtactggtc gataacatcc gctggcagtc
     5581 ctacctgtcg agcatgacca gcgccgaggc cgaggaatgg ggtgtggacg acgaccagcg
     5641 ccggttcttc gtccgcttcg gtgtgagcaa ggccaactat ggcgcaccgt tcgctgatcg
     5701 gtggttcagg cggcatgacg gcggggtgct caagcccgcc gtgctggaga ggcagcgcaa
     5761 gagcaagggg gtgccccgtg gtgaagccta agaacaagca cagcctcagc cacgtccggc
     5821 acgacccggc gcactgtctg gcccccggcc tgttccgtgc cctcaagcgg ggcgagcgca
     5881 agcgcagcaa gctggacgtg acgtatgact acggcgacgg caagcggatc gagttcagcg
     5941 gcccggagcc gctgggcgct gatgatctgc gcatcctgca agggctggtg gccatggctg
     6001 ggcctaatgg cctagtgctt ggcccggaac ccaagaccga aggcggacgg cagctccggc
     6061 tgttcctgga acccaagtgg gaggccgtca ccgctgaatg ccatgtggtc aaaggtagct
     6121 atcgggcgct ggcaaaggaa atcggggcag aggtcgatag tggtggggcg ctcaagcaca
     6181 tacaggactg catcgagcgc ctttggaagg tatccatcat cgcccagaat ggccgcaagc
     6241 ggcaggggtt tcggctgctg tcggagtacg ccagcgacga ggcggacggg cgcctgtacg
     6301 tggccctgaa ccccttgatc gcgcaggccg tcatgggtgg cggccagcat gtgcgcatca
     6361 gcatggacga ggtgcgggcg ctggacagcg aaaccgcccg cctgctgcac cagcggctgt
     6421 gtggctggat cgaccccggc aaaaccggca aggcttccat agataccttg tgcggctatg
     6481 tctggccgtc agaggccagt ggttcgacca tgcgcaagcg ccgcaagcgg gtgcgcgagg
     6541 cgttgccgga gctggtcgcg ctgggctgga cggtaaccga gttcgcggcg ggcaagtacg
     6601 acatcacccg gcccaaggcg gcaggctgac cccccccact ctattgtaaa caagacattt
     6661 ttatctttta tattcaatgg cttattttcc tgctaattgg taataccatg aaaaatacca
     6721 tgctcagaaa aggcttaaca atattttgaa aaattgccta ctgagcgctg ccgcacagct
     6781 ccataggccg ctttcctggc tttgcttcca gatgtatgct cttctgctcc cgaacgccag
     6841 caagacgtag cccagcgcgt cggccagctt gcaattcgcg ctaacttaca ttaattgcgt
     6901 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     7021 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     7081 cggtccacgt ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     7141 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     7201 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     7261 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     7321 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     7381 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     7441 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     7501 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     7561 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     7621 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     7681 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     7741 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     7801 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     7861 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     7921 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     7981 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     8041 aggttttgca ccattcgatg gtgtcaacgt aaatgccgct tcgccttcgc gcgcgaattg
     8101 caagctgatc cgggcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     8161 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     8221 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     8281 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
     8341 ataacaattt cacacaggaa acagaattca tgtcattacc acacgttatc cttactgttc
     8401 ttagcacacg cgatgcaacc gggtacgata taaccaaaga gttttctgct agcattggct
     8461 acttctggaa agccagccat cagcaggtat atcgtgaact caataaaatg ggggaacagg
     8521 gtttagtgac ttgtgttctg gaaccacagg aaggcaaacc agatcgcaag gtttattcta
     8581 taacccaagc tggccgcagc gctttaggtg agtggtttga tcaaccaacc gcacacccaa
     8641 ccgtgcgtga tgagtttagt gcgaaattga tggcttgttc agtacaatcg gcagaacctt
     8701 accgtctgca actggctgaa ctggttgaag aatcgcgtaa attggtcgct cactatcagg
     8761 aaatcgaagc cgcttattac gctaacccag ccgtgttaga caagcagcaa cgtttagagc
     8821 gtttaacgct acgtcgtaat ttactggttc gccaagcatg gattcaatgg gcggacgaag
     8881 tgttggcaga attgaacgcg atggcataaa agcttggctg ttttggcgga tgagagaaga
     8941 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     9001 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     9061 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     9121 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     9181 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     9241 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     9301 gccatcctga cggatggcct ttt
//
