LOCUS       Exported                3064 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTH18cr DNA, complete sequence.
ACCESSION   AB019609
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)
  AUTHORS   .
  TITLE     Direct Submission
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
                     /organism="Cloning vector pTH18cr"
                     /host="Escherichia coli"
                     /mol_type="other DNA"
                     /label=plasmid vector
                     /note="plasmid vector"
                     /db_xref="taxon:84732"
     CDS             1..237
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase alpha-peptide"
                     /label=lacZ
                     /note="MCS as those in pKF18c"
                     /protein_id="BAA34759.1"
                     /translation="MTMITPNSSSVPGDPLESTCRHASLLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            1..237
                     /gene="lacZ"
                     /label=lacZ
     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(534..1193)
                     /codon_start=1
                     /gene="cat-1"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat-1
                     /protein_id="BAA34760.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(534..1193)
                     /gene="cat-1"
                     /label=cat-1
     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..2857
                     /codon_start=1
                     /product="replication initiator"
                     /label=replication initiator
                     /note="wild type repA in pSC101"
                     /protein_id="BAA34761.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     protein_bind    2956..2977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."