LOCUS       V010655                 6773 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010655
VERSION     V010655
KEYWORDS    pMCSG43
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6773)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6773)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
            For ligation-independent cloning (LIC), linearize with SmaI and
            treat with T4 DNA polymerase plus dATP.
FEATURES             Location/Qualifiers
     source          1..6773
                     /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(208..225)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(232..252)
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             complement(263..283)
                     /label="TVMV site"
                     /note="tobacco vein mottling virus (TVMV) NIa protease
                     recognition and cleavage site"
     CDS             complement(305..907)
                     /gene="osmY"
                     /label="Osmotically-inducible protein Y"
                     /note="Osmotically-inducible protein Y from Escherichia
                     coli (strain K12). Accession#: P0AFH8"
     RBS             complement(915..937)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(952..976)
                     /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(977..995)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1213..1444
                     /label="dsbA promoter"
                     /note="promoter for the E. coli dsbA gene (Belin and
                     Boquet, 1994)"
     CDS             1445..2068
                     /label="DsbA"
                     /note="bacterial periplasmic oxidoreductase"
     promoter        2176..2253
                     /label="lacI promoter"
     CDS             2254..3333
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    3349..3370
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4145..4333
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4438..4580
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4766..5354)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5528..6385)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6386..6489)
                     /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 tccgttaatg atgatgatgg tggtgcccgg cggattggaa
      241 gtacaggttc tccccgggag agactctgga aacgcacggt ttccgagcct gcttttttgt
      301 acaacttagt tttcagatca tttttaacgc ttttcacacc atctaccgct ttggcgatac
      361 tttcagcacg gtcactttgt gcctgagaat cgacggtacc ggagagctga accacgccgt
      421 cggtggtttc aactttcaca tgacgggaag ggacgatatc gtccgccagc agtttggctt
      481 tgatttcact ggtggtggcg gtgtcacccg cgtagccctt caccgagcct tctttagcgt
      541 cgcgaacgtg cagtttgtcg ctgacagagg tcaccccttc aacgcctttc gccactttca
      601 ctgcctcttc ggcctgggcc tggctttcaa cgaaaccgct cagggtcacg actttttgat
      661 cggtttttac agagatatcg gtgctcttga tgttgtcatg atccaccagg gccgccttca
      721 ctttcgcggt gatggcgctg tcatccatga aattaccgac tttattcata gagctatcga
      781 ctttttgccc tgcgctttca ttggtagtct gcgcgttgtt ttccgcgtag gcagagccgg
      841 tcgcgacggc agaggtcaac attacagcca gcagagtttt cgaaatcttc agtcttgtca
      901 tagtcatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt
      961 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatcgatctc
     1021 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     1081 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     1141 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     1201 ctccttgcat gcttcgacac cgctgaaatc ggactgattg aacctttacg cgccatgcgt
     1261 ttggtttatt atcttgcctg gctaatgcgg cgttgggctg atcccgcgtt cccgaaaaat
     1321 ttcccgtggt taaccgggga agattactgg ctacgacaga cggcgacttt tatagaacag
     1381 gcaaaagttc tacaagaacc ccctttgcaa ttaacaccta tgtattaatc ggagagagta
     1441 gatcatgaaa aagatttggc tggcgctggc tggtttagtt ttagcgttta gcgcatcggc
     1501 ggcgcagtat gaagatggta aacagtacac taccctggaa aaaccggtag ctggcgcgcc
     1561 gcaagtgctg gagtttttct ctttcttctg cccgcactgc tatcagtttg aagaagttct
     1621 gcatatttct gataatgtga agaaaaaact gccggaaggc gtgaagatga ctaaatacca
     1681 cgtcaacttc atgggtggtg acctgggcaa agatctgact caggcatggg ctgtggcgat
     1741 ggcgctgggc gtggaagaca aagtcactgt tccgctgttt gaaggcgtac agaaaaccca
     1801 gaccattcgt tcagcatctg atatccgcga tgtatttatc aacgcaggta ttaaaggtga
     1861 agagtacgac gcggcgtgga acagcttcgt ggtcaaatct ctggtcgctc agcaggaaaa
     1921 agctgcagct gacgtgcaat tgcgtggcgt tccggcgatg tttgttaacg gtaaatatca
     1981 gctgaatccg cagggtatgg ataccagcaa tatggatgtt tttgttcagc agtatgctga
     2041 tacagtgaaa tatctgtccg agaaaaaata ataagcatgc accattcctt gcggcggcgg
     2101 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
     2161 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
     2221 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     2281 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     2341 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     2401 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     2461 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     2521 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     2581 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     2641 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     2701 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     2761 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     2821 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     2881 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     2941 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     3001 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     3061 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     3121 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     3181 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     3241 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     3301 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     3361 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
     3421 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     3481 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     3541 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     3601 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     3661 cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     3721 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     3781 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     3841 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     3901 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     3961 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     4021 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     4081 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     4141 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     4201 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     4261 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
     4321 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4381 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     4441 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     4501 gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa
     4561 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4621 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4681 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4741 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4801 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4861 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4921 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4981 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5041 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5101 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5161 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5221 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5281 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5341 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5401 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5461 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5521 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5581 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5641 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5701 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5761 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5821 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5881 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5941 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6001 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6061 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6121 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6181 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6241 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6301 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6361 cgacacggaa atgttgaata ctcatactct tcctttttca aattattgaa gcatttatca
     6421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6481 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tgtgaaccat
     6541 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6601 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6661 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6721 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//
