LOCUS       40924_12695        4359 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdIV DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4359)
  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 4359)
  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 4359)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4359)
  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..4359
                     /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..252
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     misc_feature    253..312
                     /gene="cspA"
                     /note="MCS = Multiple cloning sites contain the follow 
                     restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI, 
                     HindIII, SalI, PstI, XbaI"
     3'UTR           320..464
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(663..1118)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1218..1322
                     /label=AmpR promoter
     CDS             1323..2180
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2354..2942
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3080..3101)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3117..4196)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4197..4274)
                     /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 accatatgga gctcggtacc ctcgagggat ccgaattcaa gcttgtcgac
      301 ctgcagtcta gataggtaat ctctgcttaa aagcacagaa tctaagatcc ctgccatttg
      361 gcggggattt ttttatttgt tttcaggaaa taaataatcg atcgcgtaat aaaatctatt
      421 attatttttg tgaagaataa atttgggtgc aatgagaatg cgcaggccct ttcgtctcgc
      481 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      541 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      601 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      661 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      721 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      781 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      841 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      901 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      961 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1021 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1081 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     1141 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg tcaggtggca
     1201 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1261 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1321 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1381 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1441 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1501 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1561 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1621 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1681 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1741 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1801 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1861 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1921 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1981 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2041 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2101 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2161 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2221 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2281 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2341 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2401 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2461 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2521 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2581 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2641 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2701 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2761 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2821 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2881 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2941 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3001 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     3061 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3121 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3181 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     3241 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     3301 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     3361 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     3421 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     3481 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     3541 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     3601 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     3661 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     3721 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     3781 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3841 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3901 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3961 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     4021 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     4081 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     4141 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     4201 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     4261 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     4321 gcccagtagt aggttgaggc cgttgagcac cgccgccgc
//
