LOCUS       Exported                4392 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdII(SP-4) DNA, complete sequence.
ACCESSION   AB248601
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4392)
  AUTHORS   Suzuki M, Zhang J, Liu M, Woychik NA, Inouye M.
  TITLE     Single protein production in living cells facilitated by an mRNA 
            interferase
  JOURNAL   Mol. Cell 18 (2), 253-261 (2005)
  PUBMED    15837428
REFERENCE   2  (bases 1 to 4392)
  AUTHORS   Shodai T.
  TITLE     pColdI-IV(SP-4) vectors are derived from pColdI-IV(SP-2)[Ref.3], the
            ACA sequences within each transcription region were subsituted
  JOURNAL   Published Only in Database (2006)
REFERENCE   3  (bases 1 to 4392)
  AUTHORS   Shodai T.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-FEB-2006) Toshihiro Shodai, TAKARA BIO INC.; 
            Seta3-4-1, OTSU 520-2193, Japan (E-mail:shiyoudait@takara-bio.co.jp,
            Tel:81-77-543-7229, Fax:81-77-543-7238)
REFERENCE   4  (bases 1 to 4392)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4392
                     /organism="Expression vector pColdII(SP-4)"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /note="pCold vector for single-protein production (SPP) 
                     system. pColdII(SP-4) expresses an open reading frame as a 
                     fusion to a His6 containing translation enhancing element. 
                     derived from pUC118."
                     /db_xref="taxon:369923"
     gene            15..497
                     /gene="cspA"
                     /label=cspA
     regulatory      15..81
                     /regulatory_class="promoter"
                     /gene="cspA"
     promoter        15..81
                     /gene="cspA"
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     misc_feature    82..121
                     /gene="cspA"
                     /label=synthetic lac operator sequence
                     /note="synthetic lac operator sequence"
     5UTR           122..255
                     /gene="cspA"
                     /label=modified cspA 5UTR
                     /note="modified cspA 5UTR"
     misc_feature    256..270
                     /gene="cspA"
                     /label=TEE (translation enhancing element)
                     /note="TEE (translation enhancing element)"
     misc_feature    271..288
                     /gene="cspA"
                     /label=His6 tag
                     /note="His6 tag"
     CDS             271..288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    286..345
                     /gene="cspA"
                     /note="MCS = Multiple cloning sites contain the follow 
                     restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI, 
                     HindIII, SalI, PstI, XbaI"
     3UTR           353..497
                     /gene="cspA"
                     /label=cspA 3UTR
                     /note="cspA 3UTR"
     misc_feature    complement(696..1169)
                     /label=M13 intergenic region
                     /note="M13 intergenic region"
     rep_origin      complement(696..1151)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1251..1355
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1356..2216
                     /codon_start=1
                     /gene="Bla"
                     /product="beta-lactamase"
                     /label=Bla
                     /protein_id="BAE79260.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1356..2216
                     /gene="Bla"
                     /label=Bla
     rep_origin      2376..2990
                     /gene="ColE1 origin"
     gene            2376..2990
                     /gene="ColE1 origin"
                     /label=ColE1 origin
     rep_origin      2387..2975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3147..4229)
                     /codon_start=1
                     /gene="lacI"
                     /product="lactose operon repressor"
                     /label=lacI
                     /protein_id="BAE79261.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3147..4229)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(4230..4307)
                     /gene="lacI"
                     /label=lacI promoter