LOCUS       pET-32c(+).        5901 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing thioredoxin fusion proteins with an 
            enterokinase site. For other reading frames, use pET-32a(+) or 
            pET-32b(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-32c(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5901)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5901)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5901
                     /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(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    158..218
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(220..234)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(250..294)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(301..318)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(328..345)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(367..693)
                     /label=TrxA
                     /note="E. coli thioredoxin"
     RBS             complement(701..723)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(738..762)
                     /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(763..781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1094..1171
                     /label=lacI promoter
     CDS             1172..2251
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2267..2288
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3063..3251
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3356..3498
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3684..4272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4446..5303)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5304..5408)
                     /label=AmpR promoter
     rep_origin      complement(5435..5890)
                     /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 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccacagata tcccatggcc ttgtcgtcgt cgtcggtacc
      241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatacc
      301 agaaccgcgt ggcaccagac cagaagaatg atgatgatga tggtgcatat ggccagaacc
      361 agaaccggcc aggttagcgt cgaggaactc tttcaactga cctttagaca gtgcacccac
      421 tttggttgcc gccacttcac cgtttttgaa cagcagcaga gtcgggatac cacggatgcc
      481 atatttcggc gcagtgccag ggttttgatc gatgttcagt tttgcaacgg tcagtttgcc
      541 ctgatattcg tcagcgattt catccagaat cggggcgatc attttgcacg gaccgcacca
      601 ctctgcccag aaatcgacga ggatcgcccc gtccgctttg agtacatccg tgtcaaaact
      661 gtcgtcagtc aggtgaataa ttttatcgct catatgtata tctccttctt aaagttaaac
      721 aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt
      781 aatttcgcgg gatcgagatc gatctcgatc ctctacgccg gacgcatcgt ggccggcatc
      841 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
      901 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
      961 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg
     1021 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
     1081 cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     1141 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     1201 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     1261 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     1321 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     1381 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     1441 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     1501 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     1561 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     1621 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     1681 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     1741 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     1801 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     1861 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     1921 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt
     1981 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     2041 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     2101 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     2161 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     2221 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     2281 tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct
     2341 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     2401 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     2461 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     2521 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     2581 ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag
     2641 gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa
     2701 gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc
     2761 gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct
     2821 gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct
     2881 ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac
     2941 cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc
     3001 tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat
     3061 cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat
     3121 taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat
     3181 cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg
     3241 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     3301 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     3361 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     3421 gcagattgta ctgagagtgc accatatatg cggtgtgaaa taccgcacag atgcgtaagg
     3481 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4081 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4741 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5401 gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa tattttgtta
     5461 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5521 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     5581 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5641 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5701 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5761 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5821 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5881 cagggcgcgt cccattcgcc a
//
