pTH18cr vector (V002723)

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)."