LOCUS       40924_12665        4392 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdII DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4392)
  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 4392)
  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 4392)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4392)
  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..4392
                     /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"
     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"
     3'UTR           353..497
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     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
                     /label=AmpR promoter
     CDS             1356..2213
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2387..2975
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3113..3134)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3150..4229)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4230..4307)
                     /label=lacI promoter
ORIGIN
        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
      241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcatat ggagctcggt
      301 accctcgagg gatccgaatt caagcttgtc gacctgcagt ctagataggt aatctctgct
      361 taaaagcaca gaatctaaga tccctgccat ttggcgggga tttttttatt tgttttcagg
      421 aaataaataa tcgatcgcgt aataaaatct attattattt ttgtgaagaa taaatttggg
      481 tgcaatgaga atgcgcaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
      541 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
      601 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      661 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
      721 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
      781 caaaatccct tataaatcaa aagaatagcc cgagataggg ttgagtgttg ttccagtttg
      841 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
      901 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
      961 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     1021 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     1081 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     1141 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
     1201 aggagaaaat accgcatcag gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1261 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1321 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1381 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1441 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1501 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1561 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1621 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1681 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1741 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1801 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1861 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1921 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1981 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2041 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2101 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2161 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2221 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2281 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2521 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3001 ggttcctggc cttttgctgg ccttttgctc acatagtcat gccccgcgcc caccggaagg
     3061 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     3121 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3181 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     3241 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     3301 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     3361 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     3421 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     3481 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     3541 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     3601 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     3661 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     3721 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     3781 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     3841 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     3901 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     3961 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     4021 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     4081 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     4141 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     4201 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     4261 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     4321 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     4381 caccgccgcc gc
//
