LOCUS       pEX-N-His-GST.        5227 bp DNA     circular SYN 01-JAN-1980
DEFINITION  PrecisionShuttle(TM) bacterial vector for inducible expression of a 
            protein with a cleavable N-terminal 6xHis-GST tag.
ACCESSION   .
VERSION     .
KEYWORDS    pEX-N-His-GST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5227)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5227)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Inserts are typically cloned between the SgfI (AsiSI) and MluI 
            sites.
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     155..171
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        207..225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    226..250
                     /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)."
     RBS             254..276
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             284..286
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             287..304
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             311..964
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     CDS             986..1006
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    1007..1093
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    1060..1062
                     /label=stop codon
                     /note="stop codon"
     terminator      1132..1178
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1424..1501
                     /label=lacI promoter
     CDS             1502..2581
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2597..2618
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2633..2663
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2695..2711
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2727..2743)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3035..3623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3797..4654)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4655..4759)
                     /label=AmpR promoter
     rep_origin      4786..5227
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
       61 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
      121 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccgaattc
      181 tgctggactt tccaaaatgt cgagcttaat acgactcact ataggggaat tgtgagcgga
      241 taacaattcc ccttttgttt aactttaaga aggagaggcc accatgcatc accatcacca
      301 tcacgcaggc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
      361 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
      421 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
      481 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
      541 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
      601 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
      661 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
      721 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
      781 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
      841 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
      901 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
      961 aaaatcggat atcatcccaa cgaccgaaaa cctgtatttt cagggcgcga tcgccggcgc
     1021 gccagatctc aagcttaact agctagcgga ccgacgcgtt aaacgcggcc gctcgaggtt
     1081 taaacggccg gccacatgac tgactgagcc acgataaccg ctgagcaata actagcataa
     1141 ccccttgggg cctctaaacg ggtcttgagg ggttttttcc caccagcctt gtcctaatcc
     1201 gcggcggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1261 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1321 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     1381 tctccttacg catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg
     1441 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     1501 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     1561 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     1621 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     1681 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     1741 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     1801 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgagtcag
     1861 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     1921 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     1981 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     2041 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     2101 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     2161 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     2221 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     2281 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     2341 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     2401 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     2461 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     2521 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2581 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact
     2641 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     2701 acagctatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2761 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2821 aatagcgaag aggcccgcac cgatggatcc gctcttccgc ttcctcgctc actgactcgc
     2881 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2941 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3001 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3061 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3121 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3181 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3241 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3301 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3361 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3421 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3481 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3541 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3601 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3661 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3721 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3781 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3841 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3901 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3961 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4021 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4081 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4141 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4201 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4261 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4321 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4381 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4441 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4501 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4561 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4621 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4681 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4741 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     4801 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4861 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4921 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4981 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5041 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5101 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5161 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5221 aaatatt
//
