LOCUS       sequence-332370-        6544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone plasmid to pPM1001, pPM1002, pPM1003, pPM2212 and pPM2213.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6544)
  AUTHORS   Manning PA, Heuzenroeder MW, Yeadon J, Leavesley DI, Reeves PR, 
            Rowley D
  TITLE     Molecular cloning and expression in Escherichia coli K-12 of the O 
            antigens of the Inaba and Ogawa serotypes of the Vibrio cholerae O1 
            lipopolysaccharides and their potential for vaccine development.
  JOURNAL   Infect Immun. 1986 Aug;53(2):272-7. doi: 
            10.1128/IAI.53.2.272-277.1986.
  PUBMED    2426198
REFERENCE   2  (bases 1 to 6544)
  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"; doi: 
            "10.1128/IAI.53.2.272-277"; journalName: "Infect Immun"; date: 
            "1986-08"; volume: "53"; issue: "2"; pages: "272-7"
COMMENT     Sequence Label: pHC79
FEATURES             Location/Qualifiers
     source          1..6544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    138..278
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     193..212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(293..310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(464..1052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(544..563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1223..2014,2015..2083))
                     /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     1846..1865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2084..2188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2256..2274
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2301..2306,2307..2323,2324..2329)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2377..3567
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(2729..2748)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      3828..3922
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             join(6389..6544,1..36)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
      181 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
      241 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
      301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      901 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1501 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     1561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     1861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     2221 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     2281 gtcttcaaga attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta
     2341 tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc
     2401 gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg
     2461 ccgggcctct tgcgggatat cgtccattcc gacagcatcg ccagtcacta tggcgtgctg
     2521 ctagcgctat atgcgttgat gcaatttcta tgcgcacccg ttctcggagc actgtccgac
     2581 cgctttggcc gccgcccagt cctgctcgct tcgctacttg gagccactat cgactacgcg
     2641 atcatggcga ccacacccgt cctgtggatc ctctacgccg gacgcatcgt ggccggcatc
     2701 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     2761 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     2821 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg
     2881 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
     2941 cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     3001 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     3061 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     3121 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
     3181 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg
     3241 ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt
     3301 cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta
     3361 gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg
     3421 atcattggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg
     3481 ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt
     3541 gcatggagcc gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca
     3601 ccactccaag aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc
     3661 cttggcagaa catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg
     3721 gcagcgttgg gtcctggcca cgggtgcgca tgatctgcca cattacgctc ctgtccggca
     3781 aagttacctc tgccgaagtt gagtattttt gctgtatttg tcataatgac tcctgttgat
     3841 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     3901 ggcgtttttt attggtgaga atcgcagcaa cttgtcgcgc caatcgagcc atgtcgtcgt
     3961 caacgacccc ccattcaaga acagcaagca gcattgagaa ctttggaatc cagtccctct
     4021 tccacctgct gatctgcgac ttatcaacgc ccacagcttc cgctgtcttc tcagttccaa
     4081 gcattgcgat tttgttaagc aacgcactct cgattcgtag agcctcgttg cgtttgtttg
     4141 cacgaaccat atgtaagtat ttccttagat aacaattgat tgaatgtatg caaataaatg
     4201 catacaccat aggtgtggtt taatttgatg ccctttttca gggctggaat gtgtaagagc
     4261 ggggttattt atgctgttgt ttttttgtta ctcgggaagg gctttacctc ttccgcataa
     4321 acgcttccat cagcgtttat agttaaaaaa atctttcggc ctgcatgaat ggccttgttg
     4381 atcgcgcttt gatatacgcc gagatcttgg cagtcacagc atgcgcatat ccatgcttcg
     4441 accatgcgct cacaaagtag gtgaatgcgc aatgtagtac ccacatcgtc atcgctttcc
     4501 actgctctcg cgaataaaga tggaaaatca atctcatggt aatagtccat gaaaatcctt
     4561 gtattcataa atcctccagg tagctatatg caaattgaaa caaaagagat ggtgatcttt
     4621 ctaagagatg atggaatctc ccttcagtat cccgatggtc aatgcgctgg atatgggata
     4681 gatgggaata tgctgatttt tatgggacag agttgcgaac tgttcccaac taaaatcatt
     4741 ttgcacgatc agcgcactac gaactttacc cacaaatagt caggtaatga atcctgatat
     4801 aaagacaggt tgataaatca gtcttctacg cgcatcgcac gcgcacaccg tagaaagtct
     4861 ttcagttgtg agcctgggca aaccgttaac tttcggcggc tttgctgtgc gacaggctca
     4921 cgtctaaaag gaaataaatc atgggtcata aaattatcac gttgtccggc gcggcgacgg
     4981 atgttctgta tgcgctgttt ttccgtggcg cgttgctgtc tggtgatctg ccttctaaat
     5041 ctggcacagc cgaattgcgc gagcttggtt ttgctgaaac cagacacaca gcaactgaat
     5101 accagaaaga aaatcacttt acctttctga catcagaagg gcagaaattt gccgttgaac
     5161 acctggtcaa tacgcgtttt ggtgagcagc aatattgcgc ttcgatgacg cttggcgttg
     5221 agattgatac ctctgctgca caaaaggcaa tcgacgagct ggaccagcgc attcgtgaca
     5281 ccgtctcctt cgaacttatt cgcaatggag tgtcattcat caaggacgcc gctatcgcaa
     5341 atggtgctat ccacgcagcg gcaatcgaaa cacctcagcc ggtgaccaat atctacaaca
     5401 tcagccttgg tatccagcgt gatgagccag cgcagaacaa ggtaaccgtc agtgccgata
     5461 agttcaaagt taaacctggt gttgatacca acattgaaac gttgatcgaa aacgcgctga
     5521 aaaacgctgc tgaatgtgcg gcgctggatg tcacaaagca aatggcagca gacaagaaag
     5581 cgatggatga actggcttcc tatgtccgca cggccatcat gatggaatgt ttccccggtg
     5641 gtgttatctg gcagcagtgc cgtcgatagt atgcaattga taattattat catttgcggg
     5701 tcctttccgg cgatccgcct tgttacgggg cggcgacctc gcgggttttc gctatttatg
     5761 aaaattttcc ggtttaaggc gtttccgttc ttcttcgtca taacttaatg tttttattta
     5821 aaataccctc tgaaaagaaa ggaaacgaca ggtgctgaaa gcgagctttt tggcctctgt
     5881 cgtttccttt ctctgttttt gtccgtggaa tgaacaatgg aagtcaacaa aaagcagctg
     5941 gctgacattt tcggtgcgag tatccgtacc attcagaact ggcaggaaca gggaatgccc
     6001 gttctgcgag gcggtggcaa gggtaatgag gtgctttatg actctgccgc cgtcataaaa
     6061 tggtatgccg aaagggatgc tgaaattgag aacgaaaagc tgcgccggga ggttgaagaa
     6121 ctgcggcagg ccagcgaggc agatctgcat cgcaggatgc tgctggctac cctgtggaac
     6181 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     6241 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     6301 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     6361 cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat
     6421 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     6481 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     6541 ctgc
//
