LOCUS       pET-41b(+).        5932 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing GST fusion proteins with an 
            enterokinase site. For other reading frames, use pET-41a(+) or 
            pET-41c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-41b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5932)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5932)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5932
                     /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(150..173)
                     /label=8xHis
                     /note="8xHis affinity tag"
     misc_feature    174..264
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(264..278)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(309..353)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(369..386)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(396..413)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(441..1094)
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     RBS             complement(1102..1124)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1139..1163)
                     /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(1164..1182)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1495..1572
                     /label=lacI promoter
     CDS             1573..2652
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2668..2689
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3227..3415
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3520..3662
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3848..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4558..5370
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(5466..5921)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
      181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
      241 ggatccccga tatccatggg actcttgtcg tcgtcatcac cggagccacc accggtaccc
      301 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca
      361 attgcagtac taccgcgtgg caccagaccc gcggagtgat ggtgatggtg atgaccagaa
      421 ccactagttg aaccatccga ttttggagga tggtcgccac caccaaacgt ggcttgccag
      481 ccctgcaaag gccatgctat atacttgctg gatttcaagt acttatcaat ttgtgggata
      541 gcttcaatac gttttttaaa acaaactaat tttgggaacg catccaggca cattgggtcc
      601 atgtataaaa caacatcaag agcgtcatac aacatgaagt caggatgggt tacatgatca
      661 ccatttaaat atgttttatg acataaacga tcttcgaaca ttttcagcat ttcaggtagc
      721 ttgctaagaa aatcaacttt gagagtttca aagtctttac tatatgcaat tctcgaaaca
      781 ccgtatctaa tatccaaaac cgctccttca agcattgaaa tctctgcacg ctcttttgga
      841 caaccaccca acatgttgtg cttgtcagct atataacgta tgatggccat agactgtgtt
      901 aatttaacat caccatcaat ataataagga agattgggaa actccaaacc caattcaaac
      961 tttttgtttc gccatttatc accttcatcg cgctcataca aatgctcttc atatttttct
     1021 tcaagatatt ccaaaagaag tcgagtgggt tgcacaaggc ccttaatttt ccaataacct
     1081 agtatagggg acatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
     1141 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
     1201 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     1261 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     1321 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     1381 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     1441 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
     1501 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     1561 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     1621 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     1681 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     1741 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     1801 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     1861 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     1921 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     1981 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     2041 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     2101 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     2161 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     2221 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     2281 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     2341 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc
     2401 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     2461 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     2521 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     2581 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     2641 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     2701 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     2761 catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     2821 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     2881 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     2941 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
     3001 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
     3061 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
     3121 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
     3181 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
     3241 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     3301 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     3361 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     3541 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     3601 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4261 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4501 ttttggtcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     4561 agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa catggatgct
     4621 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     4681 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     4741 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     4801 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     4861 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     4921 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     4981 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     5041 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     5101 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     5161 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     5221 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     5281 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     5341 cagtttcatt tgatgctcga tgagtttttc taagaattaa ttcatgagcg gatacatatt
     5401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5461 acctgaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5521 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5581 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5641 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5701 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5761 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5821 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5881 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca
//
