LOCUS       40924_12675        4467 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdII(tet), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4467)
  AUTHORS   Schneider WM, Inouye M, Montelione GT, Roth MJ.
  TITLE     Independently inducible system of gene expression for condensed 
            single protein production (cSPP) suitable for high efficiency 
            isotope enrichment
  JOURNAL   J. Struct. Funct. Genomics 10 (3), 219-225 (2009)
  PUBMED    19642019
REFERENCE   2  (bases 1 to 4467)
  AUTHORS   Schneider WM, Roth MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-2009) Biochemistry, UMDNJ, 675 Hoes Lane, 
            Piscataway, NJ 08854, USA
REFERENCE   3  (bases 1 to 4467)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4467)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Struct. 
            Funct. Genomics"; date: "2009"; volume: "10"; issue: "3"; pages: 
            "219-225"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-MAR-2009) Biochemistry, UMDNJ, 675 Hoes Lane, Piscataway, NJ 
            08854, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(47..670)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     protein_bind    692..710
                     /label=tet operator
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    722..740
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        762..828
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     misc_feature    829..849
                     /label=tet operator 1 sequence
                     /note="tet operator 1 sequence"
     protein_bind    829..847
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     5'UTR           869..1002
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     CDS             1003..1017
                     /label=TEE
                     /note="translation enhancing element for E. coli (Qing et
                     al., 2004)"
     CDS             1018..1035
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    1033..1121
                     /note="MCS; multiple cloning site; includes NdeI, SacI,
                     BamHI, XhoI, EcoRI, HindIII, SalI, XbaI, PstI"
     3'UTR           1129..1273
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(1472..1927)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2027..2131
                     /label=AmpR promoter
     CDS             2132..2989
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3163..3751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3889..3910)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 ggttaccgtg aagttaccat cacggaaaaa ggttatgctg cttttaagac ccactttcac
       61 atttaagttg tttttctaat ccgcaaatga tcaattcaag gccgaataag aaggctggct
      121 ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa taatggcggc atactatcag
      181 tagtaggtgt ttccctttct tctttagcga cttgatgctc ttgatcttcc aatacgcaac
      241 ctaaagtaaa atgccccaca gcgctgagtg catataatgc attctctagt gaaaaacctt
      301 gttggcataa aaaggctaat tgattttcga gagtttcata ctgtttttct gtaggccgtg
      361 tacctaaatg tacttttgct ccatcgcgat gacttagtaa agcacatcta aaacttttag
      421 cgttattacg taaaaaatct tgccagcttt ccccttctaa agggcaaaag tgagtatggt
      481 gcctatctaa catctcaatg gctaaggcgt cgagcaaagc ccgcttattt tttacatgcc
      541 aatacaatgt aggctgctct acacctagct tctgggcgag tttacgggtt gttaaacctt
      601 cgattccgac ctcattaagc agctctaatg cgctgttaat cactttactt ttatctaatc
      661 tagacatcat taattcctaa tttttggctc tactctatca ttgatagagt cgcgttacca
      721 ctccctatca gtgatagaga aaagtcaaag gaatggtgtg gccgattaat cataaatatg
      781 aaaaataatt gttgcatcac ccgccaatgc gtggcttaat gcacatcaac tctatcattg
      841 atagagtttt gatgtgctag cgcatatcca gtgtagtaag gcaagtccct tcaagagtta
      901 tcgttgatac ccctcgtagt gcatattcct ttaacgcttc aaaatctgta aagcacgcca
      961 tatcgccgaa aggcgcactt aattattaag aggtaatata ccatgaatca taaagtgcat
     1021 catcatcatc atcatatgga gctcggatcc ctcgaggaat tcaagcttgt cgacctgcgt
     1081 ctagaggatc cgtcgacctg caggcatgca agctttctag ataggtaatc tctgcttaaa
     1141 agcatagaat ctaagatccc tgccatttgg cggggatttt tttatttgtt ttcaggaaat
     1201 aaataatcga tcgcgtaata aaatctatta ttatttttgt gaagaataaa tttgggtgca
     1261 atgagaatgc gcaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     1321 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1381 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     1441 tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa
     1501 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1561 atcccttata aatcaaaaga atagcccgag atagggttga gtgttgttcc agtttggaac
     1621 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1681 ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt
     1741 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1801 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     1861 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     1921 ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga
     1981 gaaaataccg catcaggcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2041 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2101 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2161 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2221 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2281 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2341 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2401 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2461 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2521 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2581 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2641 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2701 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2761 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2821 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2881 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2941 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3001 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3061 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3781 cctggccttt tgctggcctt ttgctcacat agtcatgccc cgcgcccacc ggaaggagct
     3841 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3901 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3961 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     4021 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     4081 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     4141 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     4201 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     4261 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4321 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4381 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     4441 atgggcccgc taacagcgcg atttgct
//
