LOCUS       sequence-416560-        6050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express waaK under the control of PBAD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6050)
  AUTHORS   Allard N, Collette A, Paquette J, Rodrigue S, Cote JP
  TITLE     Systematic investigation of recipient cell genetic requirements 
            reveals important surface receptors for conjugative transfer of 
            IncI2 plasmids.
  JOURNAL   Commun Biol. 2023 Nov 16;6(1):1172. doi: 10.1038/s42003-023-05534-2.
  PUBMED    37973843
REFERENCE   2  (bases 1 to 6050)
  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: "Commun 
            Biol."; date: "2023-11-16"; volume: "6(1):1172. doi"; pages: " 
            10.1038/s42003-023-05534-2"
COMMENT     Sequence Label: pWaaK
FEATURES             Location/Qualifiers
     source          1..6050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      121..207
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      299..326
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        345..436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(437..505,506..1297)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(655..674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1339..1794
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1426..1445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1636..1657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2626..2643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2760..3305)
                     /direction=LEFT
                     /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     complement(2840..2859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3381..3400
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3580..4458)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4485..4769
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4692..4711
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(6001..6018)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
       61 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
      121 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
      181 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
      241 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
      301 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
      361 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      421 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      481 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      541 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      601 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      661 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      721 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      781 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      841 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      901 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      961 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     1021 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1081 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1141 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1201 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1261 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1321 tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     1561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1681 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1741 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg
     1801 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1861 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1921 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1981 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2041 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2101 ccgcctacat acctcgctct gctaatcctg ttaccagtca ggcatttgag aagcacacgg
     2161 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     2221 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
     2281 ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc
     2341 ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     2401 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     2461 caccttgtcg ccttgcgtat aatatttgcc catggctagc ggagtgtata ctggcttact
     2521 atgttggcac tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc
     2581 accggtgcgt cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc
     2641 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
     2701 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     2761 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     2821 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     2881 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     2941 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     3001 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3061 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     3121 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     3181 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     3241 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     3301 ctcaagaaga tcatcttatt aatcagataa aatatttcta ggatcatgag cccgaagtgg
     3361 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     3421 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg tttgacagct
     3481 tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg
     3541 ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac
     3601 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
     3661 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
     3721 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
     3781 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
     3841 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
     3901 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
     3961 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
     4021 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
     4081 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
     4141 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
     4201 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
     4261 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
     4321 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     4381 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     4441 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     4501 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     4561 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
     4621 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
     4681 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
     4741 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagcg aattcgagct
     4801 ctaggaggca aaaatgatta aaaaaataat atttaccgtc acaccaatat tttctatacc
     4861 gccaaaagga gctgcagcgg ttgaaacatg gatttatcag gtcgcaaaaa gactttcgat
     4921 tccagtaact atcgcctgta ttcgaaacga aggttaccca acctacacaa aagtaaatga
     4981 tcactgtgat attcactaca ttggttttag tcgagtctat aaacgacttt tccaaaagtg
     5041 gactcgactt gatcctgtgc catattctca gcgtatttta aatacccaac ctaaaaacac
     5101 tgatccgaaa gactctgtta ttgttatcca caatagcatg aagctgtata aacagattcg
     5161 tagcagaagc cctcacgcgc agctagttat gcatcttcat aatgctttcg agccaggtga
     5221 gttagacaaa aatgcaaaga taattgttcc tagccagttt cttttcgatt tctattcgga
     5281 aaaaatgcct gatgcaaata ttgctattgt gcctaatggt ttttgttctg aatcgtatga
     5341 acaaaataat actgaagatt tgaggcagaa actgaacatt gatgcaaatg ataccgtgtt
     5401 attatttgca ggaaggatat caccagataa aggttgcctg atgcttatgg aggcttttaa
     5461 tcagctgaat aagatacaag ataacctcaa actggtgatt gtcggagatc cttttgccag
     5521 taaaaaaggc gaaaaagttg aatatcagaa aaaagttctg gatgcagcga aagctattgg
     5581 agcgcaatgc attatggcgg gtggacagcc tccagagcag atgcataact attaccgttt
     5641 agcagattta gttgttgttc cttctcaggt agaagaagct ttttgtatgg tagctgttga
     5701 agcgatggcg gcaggtaaac cagtgcttgc cagtcaaaaa ggtggaatta gtgagtttgt
     5761 tctggaagga attacaggct atcaccttgc agagcctatg acatctgaaa gtattctcgc
     5821 ggatattaag cgcgttttgg cggatgctga cagagctcaa attgctaaaa atgcccgaaa
     5881 ttttgttttc tcaaaatata gctgggaaca tgttactcag cgttttgaag cacaaataca
     5941 agactggttt ggttaagcat gcaagcttgg ctgttttggc ggatgagaga agattttcag
     6001 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
//
