LOCUS       sequence-222992-        3285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-29-1 split N-terminal T7 RNAP variant-GGSGSGSS-FRB expression 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3285)
  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 3285)
  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: p5-79
FEATURES             Location/Qualifiers
     source          1..3285
                     /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             2497..3285
                     /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 cgagcgccgt
     1981 gttatggcac gaacagtggt atgagggttt ggatgacgca tcgcgccagt tcttcggaga
     2041 acataacacc gaaaaaatgt ttgcggcact ggaacctctt tatgagatgc ttaaacgtgg
     2101 gcctgaaacg ttgcgtgaga tttcttttca gaactcattt gggcgtgatt tgaacgatgc
     2161 ctacgaatgg ctgatgaatt acaaaaagtc gaaagatgta tctaacctga atcaagcatg
     2221 ggatatttac tataatgtct ttcgtaaaat cggctaatgg agattttcaa catgctccct
     2281 caatcggttg aatgtcgccc ttttgtcttt ggcgctggta aaccatatga attttctatt
     2341 gattgtgaca aaataaactt attccgtggt gtctttgcgt ttcttttata tgttgccacc
     2401 tttatgtatg tattttctac gtttgctaac atactgcgta ataaggagtc ttaatcatgc
     2461 cagttctttt gggtattccg tagaaaaagg aagagtatga gggaagcggt gatcgccgaa
     2521 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
     2581 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt
     2641 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac
     2701 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
     2761 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
     2821 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
     2881 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
     2941 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
     3001 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
     3061 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
     3121 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
     3181 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
     3241 tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaa
//
