LOCUS       V001293                 5634 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001293
VERSION     V001293
KEYWORDS    pWUR702
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5634)
  AUTHORS   Swarts DC, Jore MM, Westra ER, Zhu Y, Janssen JH, Snijders AP, Wang
            Y, Patel DJ, Berenguer J, Brouns SJ, van der Oost J
  TITLE     DNA-guided DNA interference by a prokaryotic Argonaute.
  JOURNAL   Nature. 2014 Mar 13;507(7491):258-61. doi: 10.1038/nature12971. Epub
            2014 Feb 16.
   PUBMED   24531762
REFERENCE   2  (bases 1 to 5634)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5634)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1038/nature12971"; journalName: "Nature"; date: "2014-03-13-
            13"; volume: "507"; issue: "7491"; pages: "258-61"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             80..103
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             110..133
                     /label="HRV 3C site"
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             143..2197
                     /gene="ago"
                     /label="Protein argonaute"
                     /note="Protein argonaute from Thermus thermophilus (strain
                     ATCC BAA-163 / DSM 7039 / HB27). Accession#: Q746M7"
     CDS             2225..2242
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             2249..2275
                     /label="9xHis"
                     /note="9xHis affinity tag"
     terminator      2315..2362
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(2536..3324)
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(3325..3416)
                     /label="AmpR promoter"
     rep_origin      complement(3464..4202)
                     /direction=LEFT
                     /label="CloDF13 ori"
                     /note="Plasmids containing the CloDF13 (CDF) origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(4378..4399)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4415..5494)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(5495..5572)
                     /label="lacI promoter"
     promoter        5618..5634
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgtcaagct ggagccaccc gcagttcgaa aagggtgcac ttgaagtcct
      121 ctttcaggga cccgggtacc agatgaacca ccttggaaaa acggaagtgt tcctaaaccg
      181 cttcgcccta cggcctctga accccgagga actcaggcct tggcgcctcg aggtggttct
      241 ggaccctcct ccggggcggg aggaggtgta tccccttctc gcccaggtgg cccgccgggc
      301 ggggggcgtc acggtgcgca tgggagacgg cctcgcctcc tggtcacccc ctgaggtcct
      361 ggtcttggag ggcaccttgg cgcggatggg gcagaactac gcctaccgcc tctaccccaa
      421 ggggaggagg cctctggacc ccaaggaccc gggggagcgg agcgtgcttt cggccctagc
      481 ccgaaggctc cttcaggagc gcctcaggcg cctcgagggg gtctgggtgg aggggcttgc
      541 ggtgtaccgg agggagcacg cccgggggcc cgggtggcgg gtgcttgggg gggcggtctt
      601 ggacctttgg gtctccgact cgggggcgtt cctcctggag gtggaccccg cttaccgcat
      661 cctctgcgag atgtccctcg aggcctggct tgcccaaggc caccctctgc ccaaacgggt
      721 ccggaacgcc tacgaccggc gcacctggga gcttctccgg ctgggagagg aagaccccaa
      781 ggagctccct ctcccgggcg gcttgagcct cctggactac cacgctttca agggccgcct
      841 ccagggccgg gaaggggggc gggtggcctg ggtggcggac ccaaaggatc cgcgaaagcc
      901 aatcccccat ctgacgggcc ttttggtgcc cgttctgacc ctggaagacc ttcatgaaga
      961 ggagggaagc ctggccctct ccttgccttg ggaagagcgg cgtcggcgga cccgggagat
     1021 cgccagctgg atcggccggc gcctggggct gggcacacct gaggcggtgc gcgcccaggc
     1081 ttaccgcttg agcatcccca agctcatggg cagaagagcc gtgagcaagc ctgcggacgc
     1141 cctccgcgtg gggttttacc gggcccagga aaccgccctg gccctcttgc gcctggacgg
     1201 ggcccaagga tggccggagt ttctccggcg ggccttgctc cgggcctttg gggcgagcgg
     1261 ggcttccctc cgtttgcaca ccctccacgc ccatccctcc caaggcctgg ctttccgcga
     1321 ggcgctgcgg aaggccaagg aggagggggt ccaggccgtg ctggtcctca ccccgcccat
     1381 ggcctgggaa gaccgcaacc gcctgaaggc ccttcttctc agggaaggcc ttcccagcca
     1441 aatcctcaac gtccccctcc gggaggagga acgccaccgc tgggagaacg ccctcctggg
     1501 cctcctggcc aaagcggggc ttcaggtggt ggccctgagc ggcgcctatc cggcggagct
     1561 cgccgtgggc tttgacgccg gcggaaggga gtcctttcgc ttcgggggcg cggcctgcgc
     1621 cgtgggcggg gacggcggcc acctcctctg gaccctcccc gaggcccagg ccggggagcg
     1681 gatcccccag gaggtggtct gggacctctt ggaggagacc ctctgggcct tcagacgcaa
     1741 ggcggggagg cttccttccc gggtcctcct ccttcgggac ggccgcgtgc cccaggacga
     1801 gttcgccctg gccttggagg cccttgcccg ggaaggcata gcctacgact tggtttcggt
     1861 gcgcaagtcg ggtggggggc gggtctaccc cgtgcagggg cgcctggcgg acgggcttta
     1921 cgtccccttg gaggacaaga cctttcttct ccttaccgtc caccgggact tccggggcac
     1981 gccccgaccc ctgaagctgg tgcacgaggc gggggacacg cccctcgagg ccctggccca
     2041 ccagattttc catctgaccc gcctctaccc ggcgagcggt ttcgccttcc cccggcttcc
     2101 cgctcccctt cacctggccg accgcctggt gaaggaggtg ggccggttgg ggatccgtca
     2161 cctcaaggag gtggaccggg aaaagctctt cttcgtttag gaattcgcgg ccgcagagct
     2221 cgctctggtg ccacgcggta gttccgctca tcaccaccat catcaccatc accaccacta
     2281 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     2341 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
     2401 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     2461 ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg
     2521 aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     2581 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     2641 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     2701 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     2761 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     2821 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     2881 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     2941 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     3001 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     3061 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     3121 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     3181 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     3241 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     3301 ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca
     3361 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc
     3421 tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac
     3481 aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg
     3541 gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga
     3601 cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc
     3661 gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc
     3721 tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt
     3781 gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc gttcgctcca
     3841 agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact
     3901 gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta
     3961 actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc
     4021 aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc
     4081 acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt
     4141 tttcgtttac agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4201 tcttttctac tgaaccgctc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     4261 aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg
     4321 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     4381 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4501 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     4561 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     4621 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     4681 ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     4741 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     4801 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     4861 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     4921 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     4981 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     5041 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     5101 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     5161 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     5221 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     5281 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     5341 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     5401 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     5461 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     5521 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     5581 cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac tata
//
