LOCUS       sequence-227374-        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"
     CDS             664..675
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             942..1730
                     /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"
     primer_bind     complement(1812..1829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1946..2491)
                     /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(2026..2045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      join(3048..3093,1)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taaacaacta acggacaatt ctacctagga tccctcgaaa ggaaaaaaaa aatgaacacg
       61 attaacatcg ctaagaacga cttctctgac atcgaactgg ctgctatccc gttcaacact
      121 ctggctgacc attacggtga gcgttcagct cgcggacagt tggcccttga gcatgagtct
      181 tacgagatgg gtgaagcacg cttccgcaag atgtttgagc gtcaacttaa agctggtgag
      241 gttgcggata acgctgccgc caagcctctc atcactaccc tactccctaa gatgattgcc
      301 cgcatcaacg actggtttga ggaagtgaaa gctaagcgcg gcaggcgccc gacagccttc
      361 cagttcctaa aagaaatcaa gccggaagcc gtagcgtaca tcaccattaa gacctctctg
      421 gcttgcctaa ccagtgctga caatacaacc gttcaggctg tagcaagcgc aatcggtcgg
      481 accattgagg acgaggctcg cttcggtcgt atccgtgacc ttgaagctaa gcacttcaag
      541 aaaaacgttg aggaacaact caacaagcgc gtagggcacg tctacaaggg tggctctggc
      601 tctggctcga gcgccctcaa aaaagaattg caggcaaaca aaaaagaact tgcgcagctg
      661 aagtgggagt tacaagctct gaaaaaggaa ctggcgcagt aatggagatt ttcaacatgc
      721 tccctcaatc ggttgaatgt cgcccttttg tctttggcgc tggtaaacca tatgaatttt
      781 ctattgattg tgacaaaata aacttattcc gtggtgtctt tgcgtttctt ttatatgttg
      841 ccacctttat gtatgtattt tctacgtttg ctaacatact gcgtaataag gagtcttaat
      901 catgccagtt cttttgggta ttccgtagaa aaaggaagag tatgagggaa gcggtgatcg
      961 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     1021 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     1081 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     1141 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     1201 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     1261 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     1321 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     1381 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     1441 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     1501 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     1561 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     1621 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     1681 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa acggatcgct
     1741 tcatgtggca ggagaaaaaa gactgcaccg gtgcgtcagc agaatatgtg atacaggata
     1801 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     1861 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     1921 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     1981 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     2041 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     2101 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     2161 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     2221 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     2281 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     2341 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     2401 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     2461 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg ggtctgacgc
     2521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2821 tttatcagca ataaaccagc cagccgattc gagctcgccc ggggatcgac cagttggtga
     2881 ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat
     2941 ccttcaactc agcaaaagtt ctatttattc aacaaagccg ccgtcccgtc aagtcagcgt
     3001 aatgctctgc cagtgttaca accaattaac caattctgat ttccggtcaa ataaaacgaa
     3061 aggctcagtc gaaagactgg gcctttcgtt ttc
//
