LOCUS       pHIT198        6442 bp ds-DNA     circular SYN 18-JUN-2023
DEFINITION  pET-derived vector for expression of proteins N-terminally tagged 
            with 6xHis and maltose binding protein (multi-cloning sites: NdeI, 
            NotI, SpeI, SalI, PstI, SgfI, and XhoI)
ACCESSION   .
VERSION     .
KEYWORDS    pHIT198
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1
  AUTHORS   Aoki K, Moriguchi H, Yoshioka T, Okawa K, Tabara H
  TITLE     In vitro analyses of the production and activity of secondary small
            interfering RNAs in C. elegans
  JOURNAL   EMBO J. 2007 Dec 12;26(24):5007-19. doi: 10.1038/sj.emboj.7601910.
   PUBMED   18007599
FEATURES             Location/Qualifiers
     source          1..3073
                     /note="This plasmid is derived from pET-28a"
     regulatory      1..19
                     /regulatory_class="promoter"
                     /note="T7 RNA polymerase promoter"
     regulatory      19..46
                     /regulatory_class="operator"
                     /note="lacO"
     CDS             complement(88..1320)
                     /note="6xHis and maltose binding protein tags"
                     /codon_start=1
                     /translation="MGSSHHHHHHSSGLVPRGSHNMKTEEGKLVIWINGDKGYNGLAE
                     VGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEIT
                     PDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKE
                     LKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQP
                     SKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELA
                     KDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTHMAA
                     AMDITSRLQRSLEHHHHHH"
     misc_feature    1252..1302
                     /note="multiple cloning sites having unique restriction sites
                     (NdeI, NotI, SpeI, SalI, PstI, SgfI, and XhoI)"
     regulatory      1331..1459
                     /regulatory_class="terminator"
                     /note="T7 RNA polymerase terminator"
COMMENT     
ORIGIN
        1 taatacgact cactatagGG GAATTGTGAG CGGATAACAA TTCCCCTCTA GAAATAATTT
       61 TGTTTAACTT TAAGAAGGAG ATATACCatg GGCAGCAGCC ATCATCATCA TCATCACAGC
      121 AGCGGCCTGG TGCCGCGCGG CAGCCATAAT atgaaaactg aagaaggtaa actggtaatc
      181 tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa
      241 gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag
      301 gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc
      361 tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg
      421 tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct
      481 gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg
      541 gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc
      601 aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc
      661 aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa
      721 gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat
      781 tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg
      841 tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc
      901 ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc
      961 agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt
     1021 ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa
     1081 gagttggcga aagatccacg tattgccgcc actatggaaa acgcccagaa aggtgaaatc
     1141 atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac
     1201 gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac tCATATGGCG
     1261 GCCGCCATGG ATATCACTAG TCGACTGCAG CGATCGCTCG AGCACCACCA CCACCACCAC
     1321 TGAGATCCGG CTGCTAACAA AGCCCGAAAG GAAGCTGAGT TGGCTGCTGC CACCGCTGAG
     1381 CAATAActag cataacccct tggggcctct aaacgggtct tgaggggttt tttgCTGAAA
     1441 GGAGGAACTA TATCCGGATT GGCGAATGGG ACGCGCCCTG TAGCGGCGCA TTAAGCGCGG
     1501 CGGGTGTGGT GGTTACGCGC AGCGTGACCG CTACACTTGC CAGCGCCCTA GCGCCCGCTC
     1561 CTTTCGCTTT CTTCCCTTCC TTTCTCGCCA CGTTCGCCGG CTTTCCCCGT CAAGCTCTAA
     1621 ATCGGGGGCT CCCTTTAGGG TTCCGATTTA GTGCTTTACG GCACCTCGAC CCCAAAAAAC
     1681 TTGATTAGGG TGATGGTTCA CGTAGTGGGC CATCGCCCTG ATAGACGGTT TTTCGCCCTT
     1741 TGACGTTGGA GTCCACGTTC TTTAATAGTG GACTCTTGTT CCAAACTGGA ACAACACTCA
     1801 ACCCTATCTC GGTCTATTCT TTTGATTTAT AAGGGATTTT GCCGATTTCG GCCTATTGGT
     1861 TAAAAAATGA GCTGATTTAA CAAAAATTTA ACGCGAATTT TAACAAAATA TTAACGTTTA
     1921 CAATTTCAGG TGGCACTTTT CGGGGAAATG TGCGCGGAAC CCCTATTTGT TTATTTTTCT
     1981 AAATACATTC AAATATGTAT CCGCTCATGA ATTAATTCTT AGAAAAACTC ATCGAGCATC
     2041 AAATGAAACT GCAATTTATT CATATCAGGA TTATCAATAC CATATTTTTG AAAAAGCCGT
     2101 TTCTGTAATG AAGGAGAAAA CTCACCGAGG CAGTTCCATA GGATGGCAAG ATCCTGGTAT
     2161 CGGTCTGCGA TTCCGACTCG TCCAACATCA ATACAACCTA TTAATTTCCC CTCGTCAAAA
     2221 ATAAGGTTAT CAAGTGAGAA ATCACCATGA GTGACGACTG AATCCGGTGA GAATGGCAAA
     2281 AGTTTATGCA TTTCTTTCCA GACTTGTTCA ACAGGCCAGC CATTACGCTC GTCATCAAAA
     2341 TCACTCGCAT CAACCAAACC GTTATTCATT CGTGATTGCG CCTGAGCGAG ACGAAATACG
     2401 CGATCGCTGT TAAAAGGACA ATTACAAACA GGAATCGAAT GCAACCGGCG CAGGAACACT
     2461 GCCAGCGCAT CAACAATATT TTCACCTGAA TCAGGATATT CTTCTAATAC CTGGAATGCT
     2521 GTTTTCCCGG GGATCGCAGT GGTGAGTAAC CATGCATCAT CAGGAGTACG GATAAAATGC
     2581 TTGATGGTCG GAAGAGGCAT AAATTCCGTC AGCCAGTTTA GTCTGACCAT CTCATCTGTA
     2641 ACATCATTGG CAACGCTACC TTTGCCATGT TTCAGAAACA ACTCTGGCGC ATCGGGCTTC
     2701 CCATACAATC GATAGATTGT CGCACCTGAT TGCCCGACAT TATCGCGAGC CCATTTATAC
     2761 CCATATAAAT CAGCATCCAT GTTGGAATTT AATCGCGGCC TAGAGCAAGA CGTTTCCCGT
     2821 TGAATATGGC TCATAACACC CCTTGTATTA CTGTTTATGT AAGCAGACAG TTTTATTGTT
     2881 CATGACCAAA ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA
     2941 GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA
     3001 AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC
     3061 GAAGGTAACT GGCTTCAGCA GAGCGCAGAT ACCAAATACT GTCCTTCTAG TGTAGCCGTA
     3121 GTTAGGCCAC CACTTCAAGA ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT
     3181 GTTACCAGTG GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG
     3241 ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG
     3301 CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC
     3361 CACGCTTCCC GAAGGGAGAA AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG
     3421 AGAGCGCACG AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT
     3481 TCGCCACCTC TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG
     3541 GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA
     3601 CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC CGTATTACCG CCTTTGAGTG
     3661 AGCTGATACC GCTCGCCGCA GCCGAACGAC CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC
     3721 GGAAGAGCGC CTGATGCGGT ATTTTCTCCT TACGCATCTG TGCGGTATTT CACACCGCAT
     3781 ATATGGTGCA CTCTCAGTAC AATCTGCTCT GATGCCGCAT AGTTAAGCCA GTATACACTC
     3841 CGCTATCGCT ACGTGACTGG GTCATGGCTG CGCCCCGACA CCCGCCAACA CCCGCTGACG
     3901 CGCCCTGACG GGCTTGTCTG CTCCCGGCAT CCGCTTACAG ACAAGCTGTG ACCGTCTCCG
     3961 GGAGCTGCAT GTGTCAGAGG TTTTCACCGT CATCACCGAA ACGCGCGAGG CAGCTGCGGT
     4021 AAAGCTCATC AGCGTGGTCG TGAAGCGATT CACAGATGTC TGCCTGTTCA TCCGCGTCCA
     4081 GCTCGTTGAG TTTCTCCAGA AGCGTTAATG TCTGGCTTCT GATAAAGCGG GCCATGTTAA
     4141 GGGCGGTTTT TTCCTGTTTG GTCACTGATG CCTCCGTGTA AGGGGGATTT CTGTTCATGG
     4201 GGGTAATGAT ACCGATGAAA CGAGAGAGGA TGCTCACGAT ACGGGTTACT GATGATGAAC
     4261 ATGCCCGGTT ACTGGAACGT TGTGAGGGTA AACAACTGGC GGTATGGATG CGGCGGGACC
     4321 AGAGAAAAAT CACTCAGGGT CAATGCCAGC GCTTCGTTAA TACAGATGTA GGTGTTCCAC
     4381 AGGGTAGCCA GCAGCATCCT GCGATGCAGA TCCGGAACAT AATGGTGCAG GGCGCTGACT
     4441 TCCGCGTTTC CAGACTTTAC GAAACACGGA AACCGAAGAC CATTCATGTT GTTGCTCAGG
     4501 TCGCAGACGT TTTGCAGCAG CAGTCGCTTC ACGTTCGCTC GCGTATCGGT GATTCATTCT
     4561 GCTAACCAGT AAGGCAACCC CGCCAGCCTA GCCGGGTCCT CAACGACAGG AGCACGATCA
     4621 TGCGCACCCG TGGGGCCGCC ATGCCGGCGA TAATGGCCTG CTTCTCGCCG AAACGTTTGG
     4681 TGGCGGGACC AGTGACGAAG GCTTGAGCGA GGGCGTGCAA GATTCCGAAT ACCGCAAGCG
     4741 ACAGGCCGAT CATCGTCGCG CTCCAGCGAA AGCGGTCCTC GCCGAAAATG ACCCAGAGCG
     4801 CTGCCGGCAC CTGTCCTACG AGTTGCATGA TAAAGAAGAC AGTCATAAGT GCGGCGACGA
     4861 TAGTCATGCC CCGCGCCCAC CGGAAGGAGC TGACTGGGTT GAAGGCTCTC AAGGGCATCG
     4921 GTCGAGATCC CGGTGCCTAA TGAGTGAGCT AACTTACATT AATTGCGTTG CGCTCACTGC
     4981 CCGCTTTCCA GTCGGGAAAC CTGTCGTGCC AGCTGCATTA ATGAATCGGC CAACGCGCGG
     5041 GGAGAGGCGG TTTGCGTATT GGGCGCCAGG GTGGTTTTTC TTTTCACCAG TGAGACGGGC
     5101 AACAGCTGAT TGCCCTTCAC CGCCTGGCCC TGAGAGAGTT GCAGCAAGCG GTCCACGCTG
     5161 GTTTGCCCCA GCAGGCGAAA ATCCTGTTTG ATGGTGGTTA ACGGCGGGAT ATAACATGAG
     5221 CTGTCTTCGG TATCGTCGTA TCCCACTACC GAGATATCCG CACCAACGCG CAGCCCGGAC
     5281 TCGGTAATGG CGCGCATTGC GCCCAGCGCC ATCTGATCGT TGGCAACCAG CATCGCAGTG
     5341 GGAACGATGC CCTCATTCAG CATTTGCATG GTTTGTTGAA AACCGGACAT GGCACTCCAG
     5401 TCGCCTTCCC GTTCCGCTAT CGGCTGAATT TGATTGCGAG TGAGATATTT ATGCCAGCCA
     5461 GCCAGACGCA GACGCGCCGA GACAGAACTT AATGGGCCCG CTAACAGCGC GATTTGCTGG
     5521 TGACCCAATG CGACCAGATG CTCCACGCCC AGTCGCGTAC CGTCTTCATG GGAGAAAATA
     5581 ATACTGTTGA TGGGTGTCTG GTCAGAGACA TCAAGAAATA ACGCCGGAAC ATTAGTGCAG
     5641 GCAGCTTCCA CAGCAATGGC ATCCTGGTCA TCCAGCGGAT AGTTAATGAT CAGCCCACTG
     5701 ACGCGTTGCG CGAGAAGATT GTGCACCGCC GCTTTACAGG CTTCGACGCC GCTTCGTTCT
     5761 ACCATCGACA CCACCACGCT GGCACCCAGT TGATCGGCGC GAGATTTAAT CGCCGCGACA
     5821 ATTTGCGACG GCGCGTGCAG GGCCAGACTG GAGGTGGCAA CGCCAATCAG CAACGACTGT
     5881 TTGCCCGCCA GTTGTTGTGC CACGCGGTTG GGAATGTAAT TCAGCTCCGC CATCGCCGCT
     5941 TCCACTTTTT CCCGCGTTTT CGCAGAAACG TGGCTGGCCT GGTTCACCAC GCGGGAAACG
     6001 GTCTGATAAG AGACACCGGC ATACTCTGCG ACATCGTATA ACGTTACTGG TTTCACATTC
     6061 ACCACCCTGA ATTGACTCTC TTCCGGGCGC TATCATGCCA TACCGCGAAA GGTTTTGCGC
     6121 CATTCGATGG TGTCCGGGAT CTCGACGCTC TCCCTTATGC GACTCCTGCA TTAGGAAGCA
     6181 GCCCAGTAGT AGGTTGAGGC CGTTGAGCAC CGCCGCCGCA AGGAATGGTG CATGCAAGGA
     6241 GATGGCGCCC AACAGTCCCC CGGCCACGGG GCCTGCCACC ATACCCACGC CGAAACAAGC
     6301 GCTCATGAGC CCGAAGTGGC GAGCCCGATC TTCCCCATCG GTGATGTCGG CGATATAGGC
     6361 GCCAGCAACC GCACCTGTGG CGCCGGTGAT GCCGGCCACG ATGCGTCCGG CGTAGAGGAT
     6421 CGAGATCTCG ATCCCGCGAA AT   
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