LOCUS       40924_12720        4464 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pColdX(tet), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4464)
  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 4464)
  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 4464)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4464)
  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..4464
                     /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             1009..1026
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    1030..1118
                     /note="MCS; multiple cloning site; includes NdeI, SacI,
                     BamHI, XhoI, EcoRI, HindIII, SalI, XbaI, PstI"
     3'UTR           1126..1270
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(1469..1924)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2024..2128
                     /label=AmpR promoter
     CDS             2129..2986
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3160..3748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3886..3907)
                     /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 aggtaataca ccatgggcca tcatcatcat
     1021 catcatagcc atatggagct cggatccctc gaggaattca agcttgtcga cctgcgtcta
     1081 gaggatccgt cgacctgcag gcatgcaagc tttctagata ggtaatctct gcttaaaagc
     1141 acagaatcta agatccctgc catttggcgg ggattttttt atttgttttc aggaaataaa
     1201 taatcgatcg cgtaataaaa tctattatta tttttgtgaa gaataaattt gggtgcaatg
     1261 agaatgcgca ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacat
     1321 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     1381 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     1441 gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc
     1501 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1561 ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag
     1621 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1681 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     1741 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1801 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1861 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1921 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     1981 aataccgcat caggcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2041 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2101 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2161 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2221 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2281 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2341 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2401 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2461 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2521 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2581 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2641 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2701 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2761 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2821 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2881 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2941 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3001 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3301 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3781 ggccttttgc tggccttttg ctcacatagt catgccccgc gcccaccgga aggagctgac
     3841 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     3901 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3961 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     4021 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4081 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4141 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4201 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4261 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4321 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     4381 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     4441 ggcccgctaa cagcgcgatt tgct
//
