LOCUS       V010661                 6796 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010661
VERSION     V010661
KEYWORDS    pMCSG40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6796)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6796)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
            For ligation-independent cloning (LIC), linearize with SspI and
            treat with T4 DNA polymerase plus dGTP.
FEATURES             Location/Qualifiers
     source          1..6796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(140..157)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(223..243)
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             complement(268..285)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(286..306)
                     /label="TVMV site"
                     /note="tobacco vein mottling virus (TVMV) NIa protease
                     recognition and cleavage site"
     CDS             complement(328..930)
                     /gene="osmY"
                     /label="Osmotically-inducible protein Y"
                     /note="Osmotically-inducible protein Y from Escherichia
                     coli (strain K12). Accession#: P0AFH8"
     RBS             complement(938..960)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(975..999)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1000..1018)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1236..1467
                     /label="dsbA promoter"
                     /note="promoter for the E. coli dsbA gene (Belin and
                     Boquet, 1994)"
     CDS             1468..2091
                     /label="DsbA"
                     /note="bacterial periplasmic oxidoreductase"
     promoter        2199..2276
                     /label="lacI promoter"
     CDS             2277..3356
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    3372..3393
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4168..4356
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4461..4603
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4789..5377)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5551..6408)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6409..6512)
                     /label="AmpR promoter"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgttatcc acttccaata ttggattgga agtacaggtt
      241 ctcggtaccc agatccacgc cagaagagtg atgatgatgg tggtgagact ggaaacgcac
      301 ggtttccgag cctgcttttt tgtacaactt agttttcaga tcatttttaa cgcttttcac
      361 accatctacc gctttggcga tactttcagc acggtcactt tgtgcctgag aatcgacggt
      421 accggagagc tgaaccacgc cgtcggtggt ttcaactttc acatgacggg aagggacgat
      481 atcgtccgcc agcagtttgg ctttgatttc actggtggtg gcggtgtcac ccgcgtagcc
      541 cttcaccgag ccttctttag cgtcgcgaac gtgcagtttg tcgctgacag aggtcacccc
      601 ttcaacgcct ttcgccactt tcactgcctc ttcggcctgg gcctggcttt caacgaaacc
      661 gctcagggtc acgacttttt gatcggtttt tacagagata tcggtgctct tgatgttgtc
      721 atgatccacc agggccgcct tcactttcgc ggtgatggcg ctgtcatcca tgaaattacc
      781 gactttattc atagagctat cgactttttg ccctgcgctt tcattggtag tctgcgcgtt
      841 gttttccgcg taggcagagc cggtcgcgac ggcagaggtc aacattacag ccagcagagt
      901 tttcgaaatc ttcagtcttg tcatagtcat atgtatatct ccttcttaaa gttaaacaaa
      961 attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat
     1021 ttcgcgggat cgagatcgat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
     1081 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
     1141 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
     1201 gccgggggac tgttgggcgc catctccttg catgcttcga caccgctgaa atcggactga
     1261 ttgaaccttt acgcgccatg cgtttggttt attatcttgc ctggctaatg cggcgttggg
     1321 ctgatcccgc gttcccgaaa aatttcccgt ggttaaccgg ggaagattac tggctacgac
     1381 agacggcgac ttttatagaa caggcaaaag ttctacaaga accccctttg caattaacac
     1441 ctatgtatta atcggagaga gtagatcatg aaaaagattt ggctggcgct ggctggttta
     1501 gttttagcgt ttagcgcatc ggcggcgcag tatgaagatg gtaaacagta cactaccctg
     1561 gaaaaaccgg tagctggcgc gccgcaagtg ctggagtttt tctctttctt ctgcccgcac
     1621 tgctatcagt ttgaagaagt tctgcatatt tctgataatg tgaagaaaaa actgccggaa
     1681 ggcgtgaaga tgactaaata ccacgtcaac ttcatgggtg gtgacctggg caaagatctg
     1741 actcaggcat gggctgtggc gatggcgctg ggcgtggaag acaaagtcac tgttccgctg
     1801 tttgaaggcg tacagaaaac ccagaccatt cgttcagcat ctgatatccg cgatgtattt
     1861 atcaacgcag gtattaaagg tgaagagtac gacgcggcgt ggaacagctt cgtggtcaaa
     1921 tctctggtcg ctcagcagga aaaagctgca gctgacgtgc aattgcgtgg cgttccggcg
     1981 atgtttgtta acggtaaata tcagctgaat ccgcagggta tggataccag caatatggat
     2041 gtttttgttc agcagtatgc tgatacagtg aaatatctgt ccgagaaaaa ataataagca
     2101 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     2161 aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa
     2221 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     2281 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     2341 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     2401 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     2461 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     2521 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     2581 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     2641 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     2701 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     2761 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     2821 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     2881 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     2941 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     3001 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     3061 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     3121 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     3181 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     3241 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     3301 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3361 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc
     3421 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     3481 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     3541 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     3601 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     3661 ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg
     3721 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
     3781 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
     3841 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg
     3901 ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca
     3961 cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc
     4021 cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca
     4081 tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac
     4141 agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg
     4201 gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg
     4261 gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc
     4321 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4381 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4441 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     4501 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg
     4561 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     4621 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4681 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4741 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4801 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4861 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4921 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4981 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5041 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5101 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5161 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5221 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5281 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5341 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5401 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5461 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5521 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5581 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5641 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5701 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5761 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5821 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc
     5881 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5941 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6001 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6061 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6121 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6181 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6241 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6301 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6361 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6421 tcaaattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6481 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     6541 ttgtaagcgt taatgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6601 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6661 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6721 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6781 cgcgtcccat tcgcca
//
