LOCUS       40924_18566        3966 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETIK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3966)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Bacterial expression with pETIK
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3966)
  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 3966)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3966)
  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            "Unpublished"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..3966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        213..231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             263..285
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             305..322
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             326..358
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             362..376
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     misc_feature    376..422
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      498..545
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             692..1504
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1665..2253
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2439..2581)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2767..2844
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2845..3924
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgagatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca
      241 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgcgggg
      301 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      361 ggacgatgac gataaggatc cccgggtacc gagctcgaat tcgatttcgt cgacaagctt
      421 agcggccgcc gtttaatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      481 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      541 ttttgctgaa aggaggaact atatccggat gcgtttctac aaactctttt gtttattttt
      601 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttatgtc
      661 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
      721 ctcaaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
      781 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
      841 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
      901 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
      961 tcctgatgat gcatggttac tcaccactgc gatcccaggg aaaacagcat tccaggtatt
     1021 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1081 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1141 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1201 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagctattgc cattctcacc
     1261 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1321 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1381 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1441 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1501 tttctaatta aaacatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1561 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1621 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1681 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1741 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1801 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1861 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1921 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1981 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2041 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2101 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2161 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2221 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2281 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2341 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2401 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     2461 acgcatctgt gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg
     2521 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2581 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2641 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2701 atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt
     2761 tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     2821 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     2881 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     2941 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     3001 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     3061 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     3121 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     3181 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     3241 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     3301 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     3361 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     3421 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     3481 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     3541 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     3601 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     3661 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat
     3721 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     3781 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     3841 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3901 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt
     3961 gatctg
//
