LOCUS       sequence-333117-        7782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid pWSK29 encoding E. coli PhoA' for making reporter fusion 
            proteins for topology mapping.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7782)
  AUTHORS   Daniels C, Vindurampulle C, Morona R
  TITLE     Overexpression and topology of the Shigella flexneri O-antigen 
            polymerase (Rfc/Wzy).
  JOURNAL   Mol Microbiol. 1998 Jun;28(6):1211-22. doi: 
            10.1046/j.1365-2958.1998.00884.x.
  PUBMED    9680210
REFERENCE   2  (bases 1 to 7782)
  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: 
            "10.1046/j.1365-2958.1998.00884"; journalName: "Mol Microbiol"; 
            date: "1998-06"; volume: "28"; issue: "6"; pages: "1211-22"
COMMENT     Sequence Label: pRMCD28
FEATURES             Location/Qualifiers
     source          1..7782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1028..1978)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2026..2248
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     3393..3410
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3527..3548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3563..3568,3569..3586,3587..3593)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3601..3617
                     /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     3606..3628
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3625..3641
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3625..3641
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3660..3680
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3662..3680
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3717..3733
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3717..3733
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6156..6175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6157..6175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6185..6202)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6185..6201)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6194..6216)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6343..6798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6430..6449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6640..6661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6824..6928
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6929..6997,6998..7782,1..7)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7147..7166)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
       61 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
      121 acgtgagttt tcgttccact gagcgtcaga tctctgttcg ggggttcact gacgctgcgt
      181 gactacgatg gtcaggttgc ggaggctatg gccctggtac gagcgctgaa caaaatgacg
      241 aaagcaggta tgcctgaaag cgtgcgtatt gcctgaaaac acaacccgct acgggggaga
      301 cttacccgaa atctgattta ttcaacaaag ccactttcag tatggatttg ggtgttacgg
      361 gggggggttg gtgcttaaaa tcgcgctgag gatttctgat ggtgttaagc gggcggtttt
      421 gagatgtaaa ctcgcccgtt taacataatg gatcttgcgc gcaccgcccg aacaccactc
      481 gccacaaaaa accgccggaa cgtccaaaag tacgggtttt gctgcccgca aacgggctgt
      541 tctggtgttg ctagtttgtt atcagaatcg cagatccggc ttcagccggt ttgccggctg
      601 aaagcgctat ttcttccaga attgccatga ttttttcccc acgggaggcg tcactggctc
      661 ccgtgttgtc ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg
      721 tgactgttga gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt
      781 tttactggtt tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat
      841 ctgttcatgg tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg atgtatctat
      901 cttttttaca ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg
      961 aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat
     1021 aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt
     1081 gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt
     1141 ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta
     1201 gtccgttatg taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt
     1261 ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga
     1321 aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag
     1381 tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt
     1441 agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct
     1501 tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt
     1561 tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa
     1621 gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat
     1681 agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc
     1741 tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt
     1801 tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt
     1861 tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg
     1921 ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac
     1981 atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat
     2041 aattactagt ccttttcctt tgagttgtgg gtatctgtaa attctgctag acctttgctg
     2101 gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt
     2161 ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa
     2221 gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa
     2281 ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa
     2341 gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg
     2401 cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg
     2461 aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata
     2521 atacaagaaa agcccgtcac gggcttctca gggcgtttta tggcgggtct gcggagtggt
     2581 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     2641 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     2701 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc
     2761 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     2821 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     2881 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     2941 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     3001 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     3061 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     3121 accctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     3181 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggcattcg
     3241 ccattcaggc tgcctcctac caagacgact tcaaatccga cccgtagaga tctcacatgt
     3301 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3361 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3421 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3481 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     3541 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     3601 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     3661 caattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta
     3721 gaactagtgg atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgccc
     3781 tgttctggaa aaccgggctg ctcagggcga tattactgca cccggcggtg ctcgccgttt
     3841 aacgggtgat cagactgccg ctctgcgtga ttctcttagc gataaacctg caaaaaatat
     3901 tattttgctg attggcgatg ggatggggga ctcggaaatt actgccgcac gtaattatgc
     3961 cgaaggtgcg ggcggctttt ttaaaggtat agatgcctta ccgcttaccg ggcaatacac
     4021 tcactatgcg ctgaataaaa aaaccggcaa accggactac gtcaccgact cggctgcatc
     4081 agcaaccgcc tggtcaaccg gtgtcaaaac ctataacggc gcgctgggcg tcgatattca
     4141 cgaaaaagat cacccaacga ttctggaaat ggcaaaagcc gcaggtctgg cgaccggtaa
     4201 cgtttctacc gcagagttgc aggatgccac gcccgctgcg ctggtggcac atgtgacctc
     4261 gcgcaaatgc tacggtccga gcgcgaccag tgaaaaatgt ccgggtaacg ctctggaaaa
     4321 aggcggaaaa ggatcgatta ccgaacagct gcttaacgct cgtgccgacg ttacgcttgg
     4381 cggcggcgca aaaacctttg ctgaaacggc aaccgctggt gaatggcagg gaaaaacgct
     4441 gcgtgaacag gcacaggcgc gtggttatca gttggtgagc gatgctgcct cactgaattc
     4501 ggtgacggaa gcgaatcagc aaaaacccct gcttggcctg tttgctgacg gcaatatgcc
     4561 agtgcgctgg ctaggaccga aagcaacgta ccatggcaat atcgataagc ccgcagtcac
     4621 ctgtacgcca aatccgcaac gtaatgacag tgtaccaacc ctggcgcaga tgaccgacaa
     4681 agccattgaa ttgttgagta aaaatgagaa aggctttttc ctgcaagttg aaggtgcgtc
     4741 aatcgataaa caggatcatg ctgcgaatcc ttgtgggcaa attggcgaga cggtcgatct
     4801 cgatgaagcc gtacaacggg cgctggaatt cgctaaaaag gagggtaaca cgctggtcat
     4861 agtcaccgct gatcacgccc acgccagcca gattgttgcg ccggatacca aagctccggg
     4921 cctcacccag gcgctaaata ccaaagatgg cgcagtgatg gtgatgagtt acgggaactc
     4981 cgaagaggat tcacaagaac ataccggcag tcagttgcgt attgcggcgt atggcccgca
     5041 tgccgccaat gttgttggac tgaccgacca gaccgatctc ttctacacca tgaaagccgc
     5101 tctggggctg aaataaaacc gcgcccggca gtgaattttc gctgccgggt ggtttttttg
     5161 ctgttagcaa ccagacttaa tggcagatca cgggcgcata cgctcatggt taaaacatga
     5221 agagggatgg tgctatgaaa ataacattac tggttacctt gcttttcggt ctggtttttt
     5281 taaccaccgt cggcgctgcc gagagaactt taaccccaca acaacagcgt atgacctcct
     5341 gtaatcagca ggcgacggcg caggcgttga aaggggatgc tcgtaagacc tacatgagtg
     5401 attgcctgaa gaacagcaag tctgcgcctg gcgaaaaaag tttgacgcca cagcagcaaa
     5461 agatgcgcga atgcaataat caagcaacac aacaatctct gaaaggtgat gatcgtaata
     5521 agtttatgag tgcctgcctc aagaaagccg cctgatacct gatagtgcta acgggtgagc
     5581 tacgaaaatg gctcacccga aatatcatac ttctgccttt agctccgtct ctataatttg
     5641 ggaaaattgt ttctgaatgt tcccaaaaat aatgaatgat gaaaactttt tcaaaaaagc
     5701 ggcggcgcac ggggaggaac ctcctttaac tcctcaaaac gaacatcagc ggtccgggct
     5761 gcgcttcgcc cgtcgcgtca gactaccccg tgcggttggc ctggctggca tgttcttacc
     5821 gattgcttca acgctggttt cacacccgcc gccgggctgg tggtggctgg tgttggtcgg
     5881 ctgggcgttc gtctggccgc atttagcctg gcagatagcg agcagggccg tcgatccgct
     5941 tagccgggaa atttacaact taaaaaccga tgcagtatta gcgggaatgt gggtaggcgt
     6001 aatgggcgta aacgtgctgc cttccaccgc gatgttgatg attatgtgtc tgaatttgat
     6061 gggggcaggc ggcccccgtc tgtttgtcgc gggtctggtg ttgatggtgg tttcctgcct
     6121 tgtcaccctc gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     6181 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     6241 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     6301 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     6361 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6421 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6481 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6541 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6601 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6661 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6721 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6781 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6841 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6901 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6961 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7021 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7081 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7141 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7201 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7261 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7321 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7381 caacatgggg gatcatgtaa ctcgccttga tcgctgggaa ccggagctga atgaagccat
     7441 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7501 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7561 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7621 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7681 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7741 aaatagacag atcgctgaga taggtgcctc actgattaag ca
//
