LOCUS       40924_12660        4482 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdI(tet), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4482)
  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 4482)
  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 4482)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4482)
  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..4482
                     /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             1036..1047
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     misc_feature    1048..1136
                     /note="MCS; multiple cloning site; includes NdeI, SacI,
                     BamHI, XhoI, EcoRI, HindIII, SalI, XbaI, PstI"
     3'UTR           1144..1288
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(1487..1942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2042..2146
                     /label=AmpR promoter
     CDS             2147..3004
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3178..3766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3904..3925)
                     /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 atcatatcga aggtaggcat atggagctcg gatccctcga ggaattcaag
     1081 cttgtcgacc tgcgtctaga ggatccgtcg acctgcaggc atgcaagctt tctagatagg
     1141 taatctctgc ttaaaagcat agaatctaag atccctgcca tttggcgggg atttttttat
     1201 ttgttttcag gaaataaata atcgatcgcg taataaaatc tattattatt tttgtgaaga
     1261 ataaatttgg gtgcaatgag aatgcgcagg ccctttcgtc tcgcgcgttt cggtgatgac
     1321 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     1381 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     1441 cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt
     1501 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1561 gccgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagatagg gttgagtgtt
     1621 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1681 aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg
     1741 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1801 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1861 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1921 aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt gaaataccgc
     1981 acagatgcgt aaggagaaaa taccgcatca ggcgtcaggt ggcacttttc ggggaaatgt
     2041 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2101 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2161 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2221 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2281 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2341 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2401 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2461 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2521 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2581 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2641 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2701 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2761 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2821 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2881 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2941 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3001 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3061 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3121 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3181 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3241 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3301 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3361 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3421 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3481 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3541 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3601 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3661 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3721 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3781 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatagtca tgccccgcgc
     3841 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     3901 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4021 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4081 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4141 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4201 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4261 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4321 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4381 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     4441 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ct
//
