LOCUS       sequence-416561-        7097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express waaR and waaK under the control of PBAD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7097)
  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 7097)
  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: pWaaRK
FEATURES             Location/Qualifiers
     source          1..7097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      193..279
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      371..398
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        417..508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(509..577,578..1369)
                     /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(727..746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1411..1866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1498..1517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1708..1729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2698..2715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2832..3377)
                     /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(2912..2931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3453..3472
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3652..4530)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4557..4841
                     /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     4764..4783
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
       61 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
      121 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
      181 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      241 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
      301 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
      361 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
      421 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      481 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      541 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      601 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      661 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      721 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      781 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      841 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      901 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      961 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1021 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     1081 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1141 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1201 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1261 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1321 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1381 agtttactca tatatacttt agattgattt acgcgccctg tagcggcgca ttaagcgcgg
     1441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1621 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1741 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1801 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     1861 caatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1921 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1981 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2041 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2101 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2161 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt caggcatttg
     2221 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     2281 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt
     2341 tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca
     2401 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     2461 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     2521 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggcta gcggagtgta
     2581 tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag
     2641 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
     2701 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
     2761 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
     2821 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     2881 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     2941 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     3001 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     3061 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     3121 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     3181 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     3241 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     3301 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     3361 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc taggatcatg
     3421 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     3481 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca
     3541 tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag cagggattct
     3601 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
     3661 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     3721 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     3781 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     3841 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     3901 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     3961 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     4021 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     4081 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     4141 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     4201 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     4261 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     4321 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     4381 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     4441 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     4501 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga
     4561 aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc
     4621 gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc
     4681 catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta
     4741 tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc
     4801 tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag
     4861 cgaattcgag ctctaggagg caaaagtgaa ctcatttcct gccatagaga tagataaagt
     4921 taaagcctgg gattttaggc ttattgatga aaatactgct gaaagtttaa atgttgccta
     4981 tggtgtcgat tccaattatc ttgatggtgt aggtgtttcc attacttcaa ttgtcattaa
     5041 taatcgacat gttaatcttg atttttatat tattgttgat gtttataacg acgatttctt
     5101 tcaaaaagtt gagaaacttg ctgagcaata ccaattaaga atcacacttt atagaataaa
     5161 tactgataat cttcagtgcc tgccttgtac acaagtctgg tcaagagcaa tgtacttccg
     5221 tttatttgct tttcaattat tgggagtaac acttaatcgt ttgctttatc ttgatgccga
     5281 tgtagtttgt aagggaaata ttagtcaact attgcattta gagtttaatg gagcggtggc
     5341 tgcagtagtc agggatgtag acccaatgca ggaaaaggcg gtagtcagat tatctgatcc
     5401 tgaacttcgg gggcagtatt ttaattctgg tgtcgtttac ttagacctaa aaaaatggac
     5461 agaagcaaaa ctaaccgaaa aagcactgtc tattcttatg agtaaagata gtatttataa
     5521 gtatcctgac caggacgtta tgaatgtcct actaaaaggg atgacaatat tcctgccacg
     5581 tgagttcaat actatatata caataaaaag tgagttaaaa gataaaacgc atcaaaaata
     5641 taaagagttg ataaaagagg atactcttct tattcattat actggggcca ccaagccgtg
     5701 gcacaaatgg gcaatttatc catcggtaaa atactataaa atagctttag aaagatcccc
     5761 ctggaaagat gactcaccac gagatgcaaa atcaattatt gaattcaaaa aaagatataa
     5821 acatctgtta gtgcaacatc attatatctc aggactcatt gcaggagtgt gttatctttg
     5881 ccgaaaatat taccgtaaat aacatttaac tggtttaacg agttggctaa taatgattaa
     5941 aaaaataata tttaccgtca caccaatatt ttctataccg ccaaaaggag ctgcagcggt
     6001 tgaaacatgg atttatcagg tcgcaaaaag actttcgatt ccagtaacta tcgcctgtat
     6061 tcgaaacgaa ggttacccaa cctacacaaa agtaaatgat cactgtgata ttcactacat
     6121 tggttttagt cgagtctata aacgactttt ccaaaagtgg actcgacttg atcctgtgcc
     6181 atattctcag cgtattttaa atacccaacc taaaaacact gatccgaaag actctgttat
     6241 tgttatccac aatagcatga agctgtataa acagattcgt agcagaagcc ctcacgcgca
     6301 gctagttatg catcttcata atgctttcga gccaggtgag ttagacaaaa atgcaaagat
     6361 aattgttcct agccagtttc ttttcgattt ctattcggaa aaaatgcctg atgcaaatat
     6421 tgctattgtg cctaatggtt tttgttctga atcgtatgaa caaaataata ctgaagattt
     6481 gaggcagaaa ctgaacattg atgcaaatga taccgtgtta ttatttgcag gaaggatatc
     6541 accagataaa ggttgcctga tgcttatgga ggcttttaat cagctgaata agatacaaga
     6601 taacctcaaa ctggtgattg tcggagatcc ttttgccagt aaaaaaggcg aaaaagttga
     6661 atatcagaaa aaagttctgg atgcagcgaa agctattgga gcgcaatgca ttatggcggg
     6721 tggacagcct ccagagcaga tgcataacta ttaccgttta gcagatttag ttgttgttcc
     6781 ttctcaggta gaagaagctt tttgtatggt agctgttgaa gcgatggcgg caggtaaacc
     6841 agtgcttgcc agtcaaaaag gtggaattag tgagtttgtt ctggaaggaa ttacaggcta
     6901 tcaccttgca gagcctatga catctgaaag tattctcgcg gatattaagc gcgttttggc
     6961 ggatgctgac agagctcaaa ttgctaaaaa tgcccgaaat tttgttttct caaaatatag
     7021 ctgggaacat gttactcagc gttttgaagc acaaatacaa gactggtttg gttaagcatg
     7081 caagcttggc tgttttg
//
