LOCUS       40924_43208        3064 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTH18cr DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3064)
  AUTHORS   Hashimoto-Gotoh T, Yamaguchi M, Yasojima K, Tsujimura A, Wakabayashi
            Y, Watanabe Y.
  TITLE     A set of temperature sensitive-replication/-segregation and 
            temperature resistant plasmid vectors with different copy numbers 
            and in an isogenic background (chloramphenicol, kanamycin, lacZ, 
            repA, par, polA)
  JOURNAL   Gene 241 (1), 185-191 (2000)
  PUBMED    10607913
REFERENCE   2  (bases 1 to 3064)
  AUTHORS   Hashimoto-Gotoh T.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-NOV-1998) Contact:Tamotsu Hashimoto-Gotoh Res. Inst. 
            for Geriatrics, Kyoto Pref. Univ. Med., Biochem. and Mol. Genet.; 
            Kawaramachi-Hirokoji, Kajii-Cho 465, Kamigyo-ku, Kyoto, Kyoto 
            602-8566, Japan
REFERENCE   3  (bases 1 to 3064)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3064)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "2000"; volume: "241"; issue: "1"; pages: "185-191"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-NOV-1998) Contact:Tamotsu Hashimoto-Gotoh Res. Inst. for 
            Geriatrics, Kyoto Pref. Univ. Med., Biochem. and Mol. Genet.; 
            Kawaramachi-Hirokoji, Kajii-Cho 465, Kamigyo-ku, Kyoto, Kyoto 
            602-8566, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     On Jun 23, 1999 this sequence version replaced AB019609.1.
            The copy number of pKF38wt is 5 per chromosome. Drug concentration 
            for chloramphenicol selection should be 10 to 15 microgramms per ml.
            TOP10 but neither DH5alpha nor JM109 is advisable as a host strain 
            for alpha-complemetation selection .
            The multiple cloning sites are as in pKF18c (Gene, 1994; 152, 
            271-275).
FEATURES             Location/Qualifiers
     source          1..3064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..74
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(75..91)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(537..1193)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1194..1296)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      1637..1859
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1907..2854
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     protein_bind    2956..2977
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2992..3022
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3030..3046
                     /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)."
ORIGIN
        1 atgaccatga ttacgccgaa ttcgagctcg gtacccgggg atcctctaga gtcgacctgc
       61 aggcatgcaa gcttactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      121 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctaaccg
      241 tttttatcag gctctgggag gcagaataaa tgatcatatc gtcaattatt acctccacgg
      301 ggagagcctg agcaaactgg cctcaggcat ttgagaagca cacggtcaca ctgcttccgg
      361 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg
      421 aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact
      481 tattcaggcg tagcaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc
      541 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
      601 ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
      661 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
      721 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
      781 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
      841 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
      901 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
      961 ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     1021 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     1081 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     1141 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     1201 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     1261 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg
     1321 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     1381 tccgtcacag gtatttattc gctgtagtgc catttacccc cattcactgc cagagccgtg
     1441 agcgcagcga actgaatgtc acgaaaaaga cagcgactca ggtgcctgat ggtcggagac
     1501 aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc ctttagggtt
     1561 ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac atccttttgt aatactgcgg
     1621 aactgactaa agtagtgagt tatacacagg gctgggatct attcttttta tcttttttta
     1681 ttctttcttt attctataaa ttataaccac ttgaatataa acaaaaaaaa cacacaaagg
     1741 tctagcggaa tttacagagg gtctagcaga atttacaagt tttccagcaa aggtctagca
     1801 gaatttacag atacccacaa ctcaaaggaa aaggactagt aattatcatt gactagccca
     1861 tctcaattgg tatagtgatt aaaatcacct agaccaattg agatgtatgt ctgaattagt
     1921 tgttttcaaa gcaaatgaac tagcgattag tcgctatgac ttaacggagc atgaaaccaa
     1981 gctaatttta tgctgtgtgg cactactcaa ccccacgatt gaaaacccta caaggaaaga
     2041 acggacggta tcgttcactt ataaccaata cgctcagatg atgaacatca gtagggaaaa
     2101 tgcttatggt gtattagcta aagcaaccag agagctgatg acgagaactg tggaaatcag
     2161 gaatcctttg gttaaaggct ttgagatttt ccagtggaca aactatgcca agttctcaag
     2221 cgaaaaatta gaattagttt ttagtgaaga gatattgcct tatcttttcc agttaaaaaa
     2281 attcataaaa tataatctgg aacatgttaa gtcttttgaa aacaaatact ctatgaggat
     2341 ttatgagtgg ttattaaaag aactaacaca aaagaaaact cacaaggcaa atatagagat
     2401 tagccttgat gaatttaagt tcatgttaat gcttgaaaat aactaccatg agtttaaaag
     2461 gcttaaccaa tgggttttga aaccaataag taaagattta aacacttaca gcaatatgaa
     2521 attggtggtt gataagcgag gccgcccgac tgatacgttg attttccaag ttgaactaga
     2581 tagacaaatg gatctcgtaa ccgaacttga gaacaaccag ataaaaatga atggtgacaa
     2641 aataccaaca accattacat cagattccta cctacataac ggactaagaa aaacactaca
     2701 cgatgcttta actgcaaaaa ttcagctcac cagttttgag gcaaaatttt tgagtgacat
     2761 gcaaagtaag tatgatctca atggttcgtt ctcatggctc acgcaaaaac aacgaaccac
     2821 actagagaac atactggcta aatacggaag gatctgaggt tcttatggct cttgtatcta
     2881 tcagtgaagc atcaagacta acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa
     2941 agggaaaact gtccataatg tgagttagct cactcattag gcaccccagg ctttacactt
     3001 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3061 acat
//
