pColdI vector (V008316)

Basic Vector Information

Vector Name:
pColdI
Antibiotic Resistance:
Ampicillin
Length:
4407 bp
Type:
Expression vector
Replication origin:
ori
Source/Author:
Qing G, Ma LC, Khorchid A, Swapna GV, Mal TK, Takayama MM, Xia B, Phadtare S, Ke H, Acton T, Montelione GT, Ikura M, Inouye M.

pColdI vector Map

pColdI4407 bp600120018002400300036004200cspA promoterlac operator5'UTRTEE6xHisFactor Xa siteMCS = Multiple cloning sites contain the follow restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI, HindIII, SalI, PstI, XbaI3'UTRf1 oriAmpR promoterAmpRoriCAP binding sitelacIlacI promoter

pColdI vector Sequence

LOCUS       40924_12650        4407 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdI DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4407)
  AUTHORS   Qing G, Ma LC, Khorchid A, Swapna GV, Mal TK, Takayama MM, Xia B, 
            Phadtare S, Ke H, Acton T, Montelione GT, Ikura M, Inouye M.
  TITLE     Cold-shock induced high-yield protein production in Escherichia coli
  JOURNAL   Nat. Biotechnol. 22 (7), 877-882 (2004)
  PUBMED    15195104
REFERENCE   2  (bases 1 to 4407)
  AUTHORS   Takayama MM, Inouye M.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-AUG-2004) Masanori Mitta Takayama, UMDNJ-Robert Wood 
            Johnson Medical School, Department of Biochemistry; 675 Hoes Lane, 
            Piscataway, NJ 08854-5635, USA (E-mail:mitsutam@takara-bio.co.jp, 
            Tel:81-77-543-7200, Fax:81-77-543-2494)
REFERENCE   3  (bases 1 to 4407)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4407)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2004"; volume: "22"; issue: "7"; pages: 
            "877-882"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-AUG-2004) Masanori Mitta Takayama, UMDNJ-Robert Wood Johnson 
            Medical School, Department of Biochemistry"; volume: " 675 Hoes 
            Lane, Piscataway, NJ 08854-5635, USA 
            (E-mail:mitsutam@takara-bio.co.jp, Tel:81-77-543-7200, Fax"; pages: 
            "81-77-543-2494"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..81
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     protein_bind    84..100
                     /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)."
     5'UTR           122..255
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     CDS             256..270
                     /label=TEE
                     /note="translation enhancing element for E. coli (Qing et
                     al., 2004)"
     CDS             271..288
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             289..300
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     misc_feature    301..360
                     /gene="cspA"
                     /note="MCS = Multiple cloning sites contain the follow 
                     restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI, 
                     HindIII, SalI, PstI, XbaI"
     3'UTR           368..512
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(711..1166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1266..1370
                     /label=AmpR promoter
     CDS             1371..2228
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2402..2990
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3128..3149)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3165..4244)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4245..4322)
                     /label=lacI promoter

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