LOCUS       pET-28c(+).        5367 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expression of N-terminally 6xHis-tagged 
            proteins with a thrombin site. For other reading frames, use 
            pET-28a(+) or pET-28b(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-28c(+).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5367)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5367)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5367
                     /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..203
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(205..237)
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             complement(241..258)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(268..285)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(295..297)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(304..326)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(341..365)
                     /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(366..384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        693..770
                     /label=lacI promoter
     CDS             771..1850
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1866..1887
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2662..2850
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2955..3097
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3283..3871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3993..4805
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4901..5356)
                     /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 tccgacccat ttgctgtcca ccagtcatgc tagccatatg
      241 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt
      301 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
      361 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
      421 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
      481 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
      541 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
      601 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
      661 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
      721 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
      781 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
      841 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
      901 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
      961 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     1021 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     1081 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     1141 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     1201 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     1261 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     1321 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     1381 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     1441 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     1501 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     1561 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     1621 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     1681 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     1741 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1801 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1861 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     1921 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     1981 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     2041 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     2101 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     2161 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     2221 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     2281 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     2341 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     2401 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     2461 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     2521 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     2581 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     2641 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     2701 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     2761 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     2821 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2881 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2941 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3001 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
     3061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     3121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3721 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaaca ataaaactgt
     3961 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     4021 gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     4081 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     4141 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     4201 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     4261 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     4321 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     4381 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     4441 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     4501 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     4561 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     4621 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     4681 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     4741 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     4801 ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt
     4921 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4981 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     5041 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     5101 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5161 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5221 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     5281 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5341 ccgctacagg gcgcgtccca ttcgcca
//
