LOCUS       pET-49b(+).        5926 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing proteins with a cleavable N-terminal
            GST tag and a cleavable C-terminal S-tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET-49b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5926)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5926)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5926
                     /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(435..1088)
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     RBS             complement(1096..1118)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1133..1157)
                     /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(1158..1176)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1489..1566
                     /label=lacI promoter
     CDS             1567..2646
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2662..2683
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3221..3409
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3514..3656
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3842..4430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4552..5364
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(5460..5915)
                     /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 atggtgatga
      361 ccagaaccac tagatggagt aggagtagga ggattgttat tagaaccgcc accaccacta
      421 gttgaaccat ccgattttgg aggatggtcg ccaccaccaa acgtggcttg ccagccctgc
      481 aaaggccatg ctatatactt gctggatttc aagtacttat caatttgtgg gatagcttca
      541 atacgttttt taaaacaaac taattttggg aacgcatcca ggcacattgg gtccatgtat
      601 aaaacaacat caagagcgtc atacaacatg aagtcaggat gggttacatg atcaccattt
      661 aaatatgttt tatgacataa acgatcttcg aacattttca gcatttcagg tagcttgcta
      721 agaaaatcaa ctttgagagt ttcaaagtct ttactatatg caattctcga aacaccgtat
      781 ctaatatcca aaaccgctcc ttcaagcatt gaaatctctg cacgctcttt tggacaacca
      841 cccaacatgt tgtgcttgtc agctatataa cgtatgatgg ccatagactg tgttaattta
      901 acatcaccat caatataata aggaagattg ggaaactcca aacccaattc aaactttttg
      961 tttcgccatt tatcaccttc atcgcgctca tacaaatgct cttcatattt ttcttcaaga
     1021 tattccaaaa gaagtcgagt gggttgcaca aggcccttaa ttttccaata acctagtata
     1081 ggggacatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt
     1141 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct
     1201 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     1261 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     1321 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
     1381 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
     1441 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
     1501 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     1561 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     1621 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     1681 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     1741 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     1801 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     1861 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     1921 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     1981 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     2041 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     2101 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     2161 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     2221 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     2281 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     2341 gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac
     2401 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     2461 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     2521 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     2581 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2641 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
     2701 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2761 tagcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     2821 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     2881 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     2941 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     3001 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     3061 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     3121 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     3181 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     3241 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     3301 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     3361 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3421 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3481 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     3541 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3601 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4501 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     4561 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
     4621 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     4681 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     4741 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     4801 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
     4861 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     4921 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagt
     4981 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     5041 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     5101 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     5161 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     5221 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     5281 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     5341 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg
     5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5461 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//
