LOCUS       sequence-222993-        3093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-29-1 split N-terminal T7 RNAP variant-linker-ZA expression 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3093)
  AUTHORS   Pu J, Zinkus-Boltz J, Dickinson BC
  TITLE     Evolution of a split RNA polymerase as a versatile biosensor 
            platform.
  JOURNAL   Nat Chem Biol. 2017 Apr;13(4):432-438. doi: 10.1038/nchembio.2299. 
            Epub 2017 Feb 13.
  PUBMED    28192413
REFERENCE   2  (bases 1 to 3093)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nchembio.2299"; journalName: "Nat Chem Biol"; date: 
            "2017-04"; volume: "13"; issue: "4"; pages: "432-438"
COMMENT     Sequence Label: p5-71
FEATURES             Location/Qualifiers
     source          1..3093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..99)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(216..761)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(296..315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1318..1364
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             2027..2038
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2305..3093
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 acggatcgct tcatgtggca ggagaaaaaa gactgcaccg gtgcgtcagc agaatatgtg
       61 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
      121 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
      181 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
      241 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
      301 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
      361 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
      421 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
      481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
      541 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
      601 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
      661 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
      721 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg
      781 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      841 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      901 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      961 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1021 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1081 cggctccaga tttatcagca ataaaccagc cagccgattc gagctcgccc ggggatcgac
     1141 cagttggtga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc
     1201 gtgatctgat ccttcaactc agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc
     1261 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat ttccggtcaa
     1321 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttctaaacaa ctaacggaca
     1381 attctaccta ggatccctcg aaaggaaaaa aaaaatgaac acgattaaca tcgctaagaa
     1441 cgacttctct gacatcgaac tggctgctat cccgttcaac actctggctg accattacgg
     1501 tgagcgttca gctcgcggac agttggccct tgagcatgag tcttacgaga tgggtgaagc
     1561 acgcttccgc aagatgtttg agcgtcaact taaagctggt gaggttgcgg ataacgctgc
     1621 cgccaagcct ctcatcacta ccctactccc taagatgatt gcccgcatca acgactggtt
     1681 tgaggaagtg aaagctaagc gcggcaggcg cccgacagcc ttccagttcc taaaagaaat
     1741 caagccggaa gccgtagcgt acatcaccat taagacctct ctggcttgcc taaccagtgc
     1801 tgacaataca accgttcagg ctgtagcaag cgcaatcggt cggaccattg aggacgaggc
     1861 tcgcttcggt cgtatccgtg accttgaagc taagcacttc aagaaaaacg ttgaggaaca
     1921 actcaacaag cgcgtagggc acgtctacaa gggtggctct ggctctggct cgagcgccct
     1981 caaaaaagaa ttgcaggcaa acaaaaaaga acttgcgcag ctgaagtggg agttacaagc
     2041 tctgaaaaag gaactggcgc agtaatggag attttcaaca tgctccctca atcggttgaa
     2101 tgtcgccctt ttgtctttgg cgctggtaaa ccatatgaat tttctattga ttgtgacaaa
     2161 ataaacttat tccgtggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
     2221 ttttctacgt ttgctaacat actgcgtaat aaggagtctt aatcatgcca gttcttttgg
     2281 gtattccgta gaaaaaggaa gagtatgagg gaagcggtga tcgccgaagt atcgactcaa
     2341 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
     2401 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
     2461 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
     2521 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
     2581 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
     2641 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
     2701 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
     2761 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
     2821 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
     2881 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
     2941 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
     3001 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
     3061 gtgaaaggcg agatcaccaa ggtagtcggc aaa
//
