LOCUS       pET-47b(+).        5203 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing proteins with a cleavable N-terminal
            6xHis tag and a cleavable C-terminal S-tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET-47b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5203)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5203)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5203
                     /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(168..212)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(213..230)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     misc_feature    234..308
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(309..332)
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             complement(342..359)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(363..365)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(373..395)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(410..434)
                     /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(435..453)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        766..843
                     /label=lacI promoter
     CDS             844..1923
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1939..1960
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2498..2686
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2791..2933
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3119..3707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3829..4641
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4737..5192)
                     /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 cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc
      181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct
      241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc
      301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg gtggtgatgt
      361 gccatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
      421 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga
      481 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
      541 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
      601 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
      661 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
      721 gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg
      781 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
      841 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
      901 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
      961 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     1021 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     1081 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     1141 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     1201 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     1261 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     1321 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     1381 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     1441 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     1501 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     1561 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     1621 tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc
     1681 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     1741 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     1801 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     1861 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1921 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc
     1981 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactag
     2041 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     2101 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     2161 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     2221 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
     2281 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
     2341 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
     2401 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     2461 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     2521 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     2581 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     2641 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     2701 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2761 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     2821 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     2881 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     2941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3541 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3781 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     3841 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
     3901 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     3961 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     4021 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     4081 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
     4141 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     4201 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagtgat
     4261 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     4321 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     4381 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     4441 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     4501 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     4561 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     4621 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
     4681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
     4741 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4801 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4861 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4921 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4981 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5041 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5101 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5161 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//
