LOCUS       40924_12690        4452 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdIII(tet), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4452)
  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 4452)
  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 4452)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4452)
  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..4452
                     /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)"
     misc_feature    1018..1106
                     /note="MCS; multiple cloning site; includes NdeI, SacI,
                     BamHI, XhoI, EcoRI, HindIII, SalI, XbaI, PstI"
     3'UTR           1114..1258
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(1457..1912)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2012..2116
                     /label=AmpR promoter
     CDS             2117..2974
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3148..3736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3874..3895)
                     /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 aggcagactt aattattaag aggtaatata ccatgaatca taaagtgcat
     1021 atggagctcg gatccctcga ggaattcaag cttgtcgacc tgcgtctaga ggatccgtcg
     1081 acctgcaggc atgcaagctt tctagatagg taatctctgc ttaaaagcat agaatctaag
     1141 atccctgcca tttggcgggg atttttttat ttgttttcag gaaataaata atcgatcgcg
     1201 taataaaatc tattattatt tttgtgaaga ataaatttgg gtgcaatgag aatgcgcagg
     1261 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1321 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1381 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     1441 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     1501 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     1561 aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     1621 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     1681 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     1741 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     1801 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     1861 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     1921 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     1981 ggcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2041 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2101 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2161 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2221 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2281 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2341 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2401 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2461 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2521 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2581 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2641 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2701 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2761 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2821 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2881 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2941 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3001 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3061 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3121 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3181 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3241 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3301 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3361 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3421 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3481 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3541 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3601 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3661 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3721 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3781 gccttttgct cacatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     3841 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     3901 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3961 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4021 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4081 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4141 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     4201 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     4261 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     4321 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     4381 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     4441 gcgcgatttg ct
//
