LOCUS       40924_12710        4503 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdTEV(tet), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4503)
  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 4503)
  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 4503)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4503)
  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..4503
                     /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"
     CDS             1048..1068
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    1069..1157
                     /note="MCS; multiple cloning site; includes NdeI, SacI,
                     BamHI, XhoI, EcoRI, HindIII, SalI, XbaI, PstI"
     3'UTR           1165..1309
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(1508..1963)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2063..2167
                     /label=AmpR promoter
     CDS             2168..3025
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3199..3787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3925..3946)
                     /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 atcatagcag cggtcgtgaa aacctgtact tccagggtca tatggagctc
     1081 ggatccctcg aggaattcaa gcttgtcgac ctgcgtctag aggatccgtc gacctgcagg
     1141 catgcaagct ttctagatag gtaatctctg cttaaaagca tagaatctaa gatccctgcc
     1201 atttggcggg gattttttta tttgttttca ggaaataaat aatcgatcgc gtaataaaat
     1261 ctattattat ttttgtgaag aataaatttg ggtgcaatga gaatgcgcag gccctttcgt
     1321 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1381 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1441 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     1501 caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1561 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1621 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1681 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     1741 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     1801 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     1861 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     1921 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac
     1981 gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgtcagg
     2041 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2101 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2161 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2221 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2281 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2341 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2401 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2461 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2521 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2581 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2641 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2701 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2761 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3001 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3061 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3121 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3181 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3241 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3301 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3361 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3421 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3481 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3541 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3601 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3661 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3721 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3781 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3841 tcacatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     3901 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     3961 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4021 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4081 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4141 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4201 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     4261 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     4321 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     4381 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     4441 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     4501 gct
//
