pTH18cs1 vector (V002722)

LOCUS       40924_43213        3064 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTH18cs1 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 AB019610.1.
            The copy number of pKF38ts1 is 14 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(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     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)."