LOCUS       40924_12680        4377 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdIII DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  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 4377)
  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 4377)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4377)
  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..4377
                     /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)"
     misc_feature    271..330
                     /gene="cspA"
                     /note="MCS = Multiple cloning sites contain the follow 
                     restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI, 
                     HindIII, SalI, PstI, XbaI"
     3'UTR           338..482
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(681..1136)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1236..1340
                     /label=AmpR promoter
     CDS             1341..2198
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2372..2960
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3098..3119)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3135..4214)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4215..4292)
                     /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 catatggagc tcggtaccct cgagggatcc
      301 gaattcaagc ttgtcgacct gcagtctaga taggtaatct ctgcttaaaa gcacagaatc
      361 taagatccct gccatttggc ggggattttt ttatttgttt tcaggaaata aataatcgat
      421 cgcgtaataa aatctattat tatttttgtg aagaataaat ttgggtgcaa tgagaatgcg
      481 caggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
      541 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      601 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      661 tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
      721 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
      781 atcaaaagaa tagcccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
      841 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
      901 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
      961 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     1021 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     1081 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta
     1141 tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     1201 atcaggcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1261 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1321 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1381 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1441 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1501 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1561 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1621 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1681 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1741 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1801 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1861 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1921 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1981 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2041 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2101 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2161 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2221 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2281 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2461 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2521 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2581 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2641 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2701 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2761 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2821 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2881 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2941 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3001 gctggccttt tgctcacata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     3061 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     3121 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3181 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     3241 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     3301 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     3361 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     3421 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     3481 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     3541 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     3601 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     3661 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     3721 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     3781 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     3841 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     3901 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     3961 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     4021 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     4081 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     4141 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     4201 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     4261 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     4321 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgc
//
