LOCUS       pET-32a(+).        5900 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-32b(+) or 
            pET-32c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-32a(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5900)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5900)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5900
                     /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..217
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(219..233)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(249..293)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(300..317)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(327..344)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(366..692)
                     /label=TrxA
                     /note="E. coli thioredoxin"
     RBS             complement(700..722)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(737..761)
                     /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(762..780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1093..1170
                     /label=lacI promoter
     CDS             1171..2250
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2266..2287
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3062..3250
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3355..3497
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3683..4271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4445..5302)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5303..5407)
                     /label=AmpR promoter
     rep_origin      complement(5434..5889)
                     /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 tccgatatca gccatggcct tgtcgtcgtc gtcggtaccc
      241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca
      301 gaaccgcgtg gcaccagacc agaagaatga tgatgatgat ggtgcatatg gccagaacca
      361 gaaccggcca ggttagcgtc gaggaactct ttcaactgac ctttagacag tgcacccact
      421 ttggttgccg ccacttcacc gtttttgaac agcagcagag tcgggatacc acggatgcca
      481 tatttcggcg cagtgccagg gttttgatcg atgttcagtt ttgcaacggt cagtttgccc
      541 tgatattcgt cagcgatttc atccagaatc ggggcgatca ttttgcacgg accgcaccac
      601 tctgcccaga aatcgacgag gatcgccccg tccgctttga gtacatccgt gtcaaaactg
      661 tcgtcagtca ggtgaataat tttatcgctc atatgtatat ctccttctta aagttaaaca
      721 aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta
      781 atttcgcggg atcgagatcg atctcgatcc tctacgccgg acgcatcgtg gccggcatca
      841 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
      901 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
      961 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
     1021 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
     1081 gtcgagatcc cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
     1141 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     1201 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     1261 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     1321 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     1381 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     1441 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     1501 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     1561 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     1621 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     1681 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     1741 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     1801 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     1861 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     1921 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga catctcggta
     1981 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     2041 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     2101 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     2161 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2221 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     2281 cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2341 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2401 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
     2461 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
     2521 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
     2581 cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     2641 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     2701 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     2761 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
     2821 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     2881 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     2941 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     3001 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     3061 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     3121 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     3181 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg
     3241 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     3301 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     3361 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     3421 cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga
     3481 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4081 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4261 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4321 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4381 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4441 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4501 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4561 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4621 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4681 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4741 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4801 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4861 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4921 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4981 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5041 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5101 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5161 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5221 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5281 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5341 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5401 ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat attttgttaa
     5461 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5521 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5581 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5641 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5701 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5761 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5821 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5881 agggcgcgtc ccattcgcca
//
