LOCUS       40924_18496        3223 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETBK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3223)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Bacterial expression with pETBK
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3223)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2009) BioDynamics Laboratory Inc., 9-7 Hongo 
            2-Chome, Bunkyo-Ku, Tokyo 113-0033, Japan
REFERENCE   3  (bases 1 to 3223)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3223)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUL-2009) BioDynamics Laboratory Inc., 9-7 Hongo 2-Chome, 
            Bunkyo-Ku, Tokyo 113-0033, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        171..189
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             221..243
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             263..280
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             284..316
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             320..334
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     misc_feature    334..380
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      456..503
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             703..1515
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1676..2264
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2450..2592)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2697..2885)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
       61 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
      121 gcacccgtgg ccaggaccca acgctgcccg agatctcgat cccgcgaaat taatacgact
      181 cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg
      241 agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg
      301 tggacagcaa atgggtcggg acgatgacga taaggatccc cgggtaccga gctcgaattc
      361 gatttcgtcg acaagcttag cggccgccgt ttaatccggc tgctaacaaa gcccgaaagg
      421 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      481 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa tgtcatgata
      541 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      661 atgcttatgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
      721 gaaacgtctt gctcaaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
      781 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
      841 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
      901 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
      961 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg gaaaacagca
     1021 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1081 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1141 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1201 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagctattg
     1261 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     1321 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     1381 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     1441 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     1501 ctcgatgagt ttttctaatt aaaacatata tactttagat tgatttaaaa cttcattttt
     1561 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1621 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1681 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1741 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1801 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1861 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1921 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1981 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2041 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2101 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2161 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2221 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2281 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2341 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2401 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     2461 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac
     2521 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2581 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2641 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2701 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     2761 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     2821 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     2881 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     2941 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     3001 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     3061 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     3121 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     3181 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcg
//
