LOCUS       40924_46568        6811 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWKS130, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  AUTHORS   Wang RF, Kushner SR.
  TITLE     Construction of versatile low-copy-number vectors for cloning, 
            sequencing and gene expression in Escherichia coli
  JOURNAL   Gene 100, 195-199 (1991)
  PUBMED    2055470
REFERENCE   2  (bases 1 to 6811)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Low copy expression vectors for use in Yersinia sp. and related 
            organisms
  JOURNAL   Plasmid 68 (1), 33-42 (2012)
  PUBMED    22445322
REFERENCE   3  (bases 1 to 6811)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2011) Microbiology and Immunology, University of 
            North Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen 
            Jones, CB# 7290, Chapel Hill, NC 27599, USA
REFERENCE   4  (bases 1 to 6811)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6811)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene 100, 
            195-199 (1991)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2012"; volume: "68"; issue: "1"; pages: "33-42"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (14-DEC-2011) Microbiology and Immunology, University of North 
            Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen Jones, 
            CB# 7290, Chapel Hill, NC 27599, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method :: DNASTAR Lasergene SeqMan Pro v. 7.2.1 (1) Intel 
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    240..261
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        276..306
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    314..330
                     /label=lac operator
                     /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     338..354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        375..393
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    complement(406..513)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(522..540)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(550..566)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      708..1136
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2454..3266)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4606..5553)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(5601..5823)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 agatctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
       61 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      121 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
      181 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      241 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct cccggctcgt
      301 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
      361 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctgggtac cgggcccccc
      421 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact
      481 agttctagag cggccgccac cgcggtggag ctccaattcg ccctatagtg agtcgtatta
      541 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      601 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      661 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
      721 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
      781 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
      841 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
      901 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
      961 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1021 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1081 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1141 caaaaaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     1201 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     1261 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     1321 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     1381 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1441 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     1501 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     1561 gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     1621 aacgcctggt tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa
     1681 agcaaattcg acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat
     1741 tgctggttta ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc
     1801 tgaggccagt ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat
     1861 tctgcctccc agagcctgat aaaaacggtt agcgccgggg ttggattttt cagcgttcca
     1921 gctaaggcta aggcattcct gttcaagcgc aagccgggct ataaagcgca ttatcgcttt
     1981 acccgtgccc ttatttcgat cgcactgaga aacatacagc tctttaatat gcaggtaatc
     2041 cgggcagcgc aacggaacat tcatcagtgt aaaaatggaa tcaataaagc cctgcgcagc
     2101 gcgcagggtc agcctgaata cgcgtttaat gaccagcaca gtcgtgatgg caaggtcaga
     2161 atagcgctga ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc
     2221 cccatcatcc agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga
     2281 ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg
     2341 cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt
     2401 caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac
     2461 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     2521 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     2581 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     2641 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     2701 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2761 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     2821 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2881 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     2941 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     3001 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     3061 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     3121 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     3181 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa
     3241 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     3301 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     3361 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     3421 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     3481 tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg
     3541 gcgattcagg cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgctat
     3601 tctgaccttg ccatcacgac tgtgctggtc attaaacgcg tattcaggct gaccctgcgc
     3661 gctgcgcagg gctttattga ttccattttt acactgatga atgttccgtt gcgctgcccg
     3721 gattacagct gttcgggggt tcactgacgc tgcgtgacta cgatggtcag gttgcggagg
     3781 ctatggccct ggtacgagcg ctgaacaaaa tgacgaaagc aggtatgcct gaaagcgtgc
     3841 gtattgcctg aaaacacaac ccgctacggg ggagacttac ccgaaatctg atttattcaa
     3901 caaagccact ttcagtatgg atttgggtgt tacggggggg ggttggtgct taaaatcgcg
     3961 ctgaggattt ctgatggtgt taagcgggcg gttttgagat gtaaactcgc ccgtttaaca
     4021 taatggatct tgcgcgcacc gcccgaacac cactcgccac aaaaaaccgc cggaacgtcc
     4081 aaaagtacgg gttttgctgc ccgcaaacgg gctgttctgg tgttgctagt ttgttatcag
     4141 aatcgcagat ccggcttcag ccggtttgcc ggctgaaagc gctatttctt ccagaattgc
     4201 catgattttt tccccacggg aggcgtcact ggctcccgtg ttgtcggcag ctttgattcg
     4261 ataagcagca tcgcctgttt caggctgtct atgtgtgact gttgagctgt aacaagttgt
     4321 ctcaggtgtt caatttcatg ttctagttgc tttgttttac tggtttcacc tgttctatta
     4381 ggtgttacat gctgttcatc tgttacattg tcgatctgtt catggtgaac agctttgaat
     4441 gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg
     4501 catatggaca gttttccctt tgatatgtaa cggtgaacag ttgttctact tttgtttgtt
     4561 agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt
     4621 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
     4681 gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
     4741 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga
     4801 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
     4861 tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc
     4921 tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt
     4981 caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa
     5041 cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc
     5101 ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc
     5161 cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa
     5221 aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa
     5281 gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt
     5341 agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt attggttata
     5401 agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc
     5461 cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag
     5521 ttcatttgct ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta
     5581 atcactatac caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt
     5641 tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc
     5701 ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat
     5761 ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa
     5821 ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc
     5881 tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa
     5941 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
     6001 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
     6061 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
     6121 tctcagggcg ttttatggcg ggtctgcgga gtggtgaatc cgttagcgag gtgccgccgg
     6181 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     6241 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
     6301 cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga
     6361 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
     6421 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
     6481 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     6541 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     6601 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     6661 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     6721 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     6781 ctgggttgaa ggctctcaag ggcatcggtc g
//
