LOCUS       sequence-456795-        5644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid induces natural competence in classical biotype Vibrio 
            cholerae and expresses the genes tfox and qstR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5644)
  AUTHORS   Woldetsadik YA, Lazinski DW, Camilli A
  TITLE     A Vibrio cholerae anti-phage system depletes nicotinamide adenine 
            dinucleotide to restrict virulent bacteriophages.
  JOURNAL   mBio. 2024 Oct 8:e0245724. doi: 10.1128/mbio.02457-24.
  PUBMED    39377576
REFERENCE   2  (bases 1 to 5644)
  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. 2024 
            Oct 8:e0245724. doi: 10.1128/mbio.02457-24."
COMMENT     Sequence Label: pDL1403
FEATURES             Location/Qualifiers
     source          1..5644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1343..1360)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1513..1599
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1691..1718
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1737..1828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1829..1897,1898..2689)
                     /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(2047..2066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2708..3253
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3154..3173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3370..3387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            3630..3739
                     /note="incP origin of transfer"
     CDS             3772..4143
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     regulatory      4155..4164
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4166..5248)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5210..5229
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5249..5326)
                     /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(5556..5561,5562..5577,5578..5584)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5592..5608
                     /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     5597..5619
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5616..5632
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5616..5632
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga taggaggtaa catgaatgag caacagtttt
       61 tcgactacgt aactaagttt ggcgcctacc aaaaacgctc gatgtttggt ggtattggct
      121 tgtttcaaca cgacgctatg tatgtgttgg tcagtgaaga ccgcattttt gtgcgtggtg
      181 gagaagagct cgaccctgag ctcttggcct tagggtgcga gaaatatcgc catgttaaaa
      241 aacagacaac ggcaaccgta aactattacg acatcactga actgtatgag caacatcacc
      301 ctgagctaga ctcgattatt gaacgctcga ttcagttttc tgtgaatcag cgggaatttc
      361 aacgctcagc agccagtcgt cgtctacggg atttacccaa tatgcaactg actttggagc
      421 gtatggtaaa aaaagcgggc attgacgatg tggaaacttt tatgagcctc ggtgcgccag
      481 aagtgtttaa taaggtgcgt caagcttatg gaagtgatgt cgatgtaaaa ctactgtgga
      541 agtttgcggg tgcgattgaa ggcattcact ggaaactgct gcaagagccg cgtaagcgcc
      601 aactgttaga aagttgtcag cagcgttagg gggtcatcat gcaacgagcc aactatgcac
      661 gcacgattta tttattaacg acacagccaa aagcgttaca tcccagtatt caagccgcca
      721 ttgagcagtt gaatctgccc gtcccggtga ttgaaccaga acgcttattg cgcgagtatc
      781 aatccgataa acacaaaata ttgttgctcg accatgcgga aaatggcctg attcgccagc
      841 aattgggtcc cctaaagctt accagcccgt acttcgaaac catcttgttt aatgtcgaga
      901 aacggttaag aacagaagat ctgctcactt ttggcaacct caaaggtctg ttttatgcca
      961 atgaagacac aggctttatt gcccacggtc ttggggagat cattaatggc caaaactggc
     1021 tacctcgcca tgtcagcagt cagttgctgc attactatcg ttacgctttc caaagccatc
     1081 aagttcaagc cacggtcgat ttgactgcac gtgaaattca gatcctgcgc tgtttacaaa
     1141 ctggcgcatc gaatatgcaa atcgctgaga atttatttat tagtgagttc acggtgaaat
     1201 cgcatcttta tcagatattt aaaaagctaa acgtgaaaaa tcgagtcaaa gcgatcgctt
     1261 gggttaatca aaatttactg taaaggcgct caacctagcg gagtgaattc ggctgttttg
     1321 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     1381 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     1441 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     1501 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1561 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     1621 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     1681 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
     1741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1801 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1861 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1921 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1981 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2041 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     2101 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2161 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2221 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2281 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2341 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2401 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2461 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2521 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2581 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2641 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaag atctccggat
     2701 gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg
     2761 ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact
     2821 ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg
     2881 acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca
     2941 tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga
     3001 acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc
     3061 aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc
     3121 aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc
     3181 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     3241 gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg
     3301 gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg
     3361 accgagcgta gcgagtcagt gagcgaggaa gcggaatgca tacgcgtctc tcgcctgtcc
     3421 cctcagttca gtaatttcct gcatttgcct gtttccagtc ggtagatatt ccacaaaaca
     3481 gcagggaagc agcgcttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt
     3541 tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac
     3601 tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg
     3661 agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct
     3721 gctctgcgag gctggccggc taccgccggc gtaacagatg agggcaagcg gatggctgat
     3781 gaaaccaagc caaccaggaa gggcagccca cctatcaagg tgtactgcct tccagacgaa
     3841 cgaagagcga ttgaggaaaa ggcggcggcg gccggcatga gcctgtcggc ctacctgctg
     3901 gccgtcggcc agggctacaa aatcacgggc gtcgtggact atgagcacgt ccgcgagctg
     3961 gcccgcatca atggcgacct gggccgcctg ggcggcctgc tgaaactctg gctcaccgac
     4021 gacccgcgca cggcgcggtt cggtgatgcc acgatcctcg ccctgctggc gaagatcgaa
     4081 gagaagcagg acgagcttgg caaggtcatg atgggcgtgg tccgcccgag ggcagagcca
     4141 tgactttttt agcggccgcc atggctcact gcccgctttc cagtcgggaa acctgtcgtg
     4201 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     4261 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     4321 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     4381 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     4441 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     4501 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     4561 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     4621 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     4681 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     4741 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     4801 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     4861 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     4921 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     4981 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     5041 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     5101 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     5161 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     5221 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     5281 gctatcatgc cataccgcga aaggttttgc accattcgat ggtgtcaacg taaatgccgc
     5341 ttcgccttcg cgcgcgaatt gcaagctgat ccgggcttat cgactgcacg gtgcaccaat
     5401 gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact
     5461 gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat
     5521 cataacggtt ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat
     5581 aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     5641 cgcc
//
