LOCUS       sequence-416565-        6982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express waaT and waaW from E. coli R4 under the control of PBAD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6982)
  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 6982)
  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: pWaaTW-R4
FEATURES             Location/Qualifiers
     source          1..6982
                     /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 caaaaatgaa tgaatttata aaagaacggt tttcgtattt
     4921 agccgataat aaaaaagaaa acgccccaga gctaaatgtt tcctacggta tcgataagaa
     4981 ttttttgtat ggtgctggcg tttcaatttc ttccgttttg attaataatt cagatattaa
     5041 ttttgtcttt catgttttca ctgattatgt ggatgatgat tatttaaagt catttaatga
     5101 aacagcaaaa caatttaata cctcaattat tgtatattta attgacccca aatactttgc
     5161 tgatctgccg acgtcacagt tttggtcgta cgcgacatac ttcagggtat tgtcttttga
     5221 atatctgagt gaaagtattt ccacactgct gtatctggat gccgatgttg tttgtaaagg
     5281 aagcctgaaa cctctcacag aaattatatt taaagatgag tttgctgcgg tcgttcctga
     5341 caatgatagt actcaggcgg catgtgcaaa acgtcttaac attcccgaaa tgaatggacg
     5401 ttatttcaat gcaggcgtta tctatgtcaa tcttaaaaaa tggcatgaag caaatttgac
     5461 accgtattta ctcacgcttt tacgaggtga aactaaatat ggctctctta aatatttaga
     5521 tcaggatgcg ttgaatatcg catttaatat gaataatatc taccttgcga aggattttga
     5581 tactatttat accctgaaaa acgaacttta tgatcgtagt catcgaaagt atcagcaaac
     5641 cattaccgat aaaacagtgt tgattcacta tacagggata actaaaccat ggcatagctg
     5701 ggctggatat ccgtctgcat catactttaa tatcgcgcgt gaacaatctc cctggaagaa
     5761 atatcctctt aaagaggcgc ggactgttgc agaaatgcag aaacaatata agcatctgtt
     5821 tgcccatggt gagtatataa aaggcataac ttcattaatt aagtacaagc ttaagaaata
     5881 aaccacacag tcaaaacgga accaacatgg atttattagc tgagagtatt actgaagtcg
     5941 ctgtctctgg ggaaattgct aacaccgatc gtgtgttaaa tatcgcttac ggtattgacc
     6001 gtaatttttt atttggtgcg gccgtatcta tgcaatcagt tgttatgcat aacccggacc
     6061 ttgcggttaa gtttcatctc tttaccgact acattgatga agattatcta caacgtgtta
     6121 acgcttttac cagcaaaaat gctaacgttg aagtaagaat ttataaagtc tccagtgcct
     6181 ttattgatat tttccccagc ctgaaacagt ggtcttatgc aacattcttc cgtttagttg
     6241 cgttccagta tctgagtgaa actattgaaa atctgttata tatcgatgct gatgtcatct
     6301 gtaaaggctc attagctgga ttgcttgata tcaattttga tggcgataag ttcgcagctg
     6361 ttattaaaga tgtgcctttt atgcaggaaa aaccagcgaa gcgtctggct atagagggac
     6421 ttccaggtaa ttatttcaac gccggtgtag tatatctgca gcttgaagca tgggcgaaaa
     6481 atgattttat gaataaagcc attgctatgc tggcaagtga cccgcagcat acgaagtata
     6541 aatgccttga tcaggatatt ttaaatattc tgttctttgg tcattgtatt tttattagcg
     6601 gcgattatga ttgcttttat ggcattgact atgagttaaa aaataaaagc gatgaagatt
     6661 ataaaaagac cattaccgat gatactaagc tgattcatta tgttggcgta acgaagccct
     6721 ggaacgactg gacgaattat ccctgccaga agtattttaa tgaggcttat caggcttctt
     6781 gctggaatga tgtggcgttt attccagcca cgaatgaaaa gcagtatcaa gtgaaatcca
     6841 gacatttgaa aagaaatggt aatattgcat cttcatttta ctatttcatg ctctattata
     6901 gtaagaaaat agcgcgtaag catttttcca aataattatt aagattaatc acccagagat
     6961 gcatgcaagc ttggctgttt tg
//
