LOCUS       pET-48b(+).        5605 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing proteins with a cleavable N-terminal
            thioredoxin tag and a cleavable C-terminal S-tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET-48b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5605)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5605)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5605
                     /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(441..767)
                     /label=TrxA
                     /note="E. coli thioredoxin"
     RBS             complement(775..797)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(812..836)
                     /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(837..855)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1168..1245
                     /label=lacI promoter
     CDS             1246..2325
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2341..2362
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2900..3088
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3193..3335
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3521..4109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4231..5043
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(5139..5594)
                     /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 gtatggccag aaccagaacc ggccaggtta gcgtcgagga actctttcaa ctgaccttta
      481 gacagtgcac ccactttggt tgccgccact tcaccgtttt tgaacagcag cagagtcggg
      541 ataccacgga tgccatattt cggcgcagtg ccagggtttt gatcgatgtt cagttttgca
      601 acggtcagtt tgccctgata ttcgtcagcg atttcatcca gaatcggggc gatcattttg
      661 cacggaccgc accactctgc ccagaaatcg acgaggatcg ccccgtccgc tttgagtaca
      721 tccgtgtcaa aactgtcgtc agtcaggtga ataattttat cgctcatatg tatatctcct
      781 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
      841 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
      901 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
      961 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     1021 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     1081 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     1141 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
     1201 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     1261 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     1321 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     1381 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     1441 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     1501 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     1561 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     1621 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     1681 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     1741 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     1801 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     1861 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     1921 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     1981 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     2041 gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     2101 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     2161 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     2221 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2281 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2341 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     2401 caacccagtc agctccttcc ggtgggcgcg gggcatgact agcatgatcg tgctcctgtc
     2461 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
     2521 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
     2581 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
     2641 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
     2701 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
     2761 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
     2821 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
     2881 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
     2941 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
     3001 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
     3061 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3121 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3181 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     3241 aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac
     3301 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3961 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat aaaactgtct
     4201 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     4261 tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     4321 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     4381 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     4441 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     4501 cctgatgatg catggttact caccactgcg atccccggca aaacagcatt ccaggtatta
     4561 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     4621 ttgcattcga ttcctgtttg taattgtcct tttaacagtg atcgcgtatt tcgtctcgct
     4681 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     4741 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     4801 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     4861 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     4921 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     4981 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     5041 ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5101 aggggttccg cgcacatttc cccgaaaagt gccacctgaa attgtaaacg ttaatatttt
     5161 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5221 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5281 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5341 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5401 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5461 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5521 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5581 gctacagggc gcgtcccatt cgcca
//
