LOCUS       sequence-454024-        3602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses WT NarX capable of sensing nitrate. For use with NarL-YdfI
            and reporter plasmids from this study..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3602)
  AUTHORS   Peruzzi JA, Galvez NR, Kamat NP
  TITLE     Engineering transmembrane signal transduction in synthetic membranes
            using two-component systems.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 May 9;120(19):e2218610120. doi: 
            10.1073/pnas.2218610120. Epub 2023 May 1.
  PUBMED    37126679
REFERENCE   2  (bases 1 to 3602)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2023-05-9"; volume: "120(19):e2218610120. 
            doi"; pages: " 10.1073/pnas.2218610120. Epub 2023 May 1"
COMMENT     Sequence Label: pJL NarX Myc
FEATURES             Location/Qualifiers
     source          1..3602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             683..705
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2519..2548
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2603..2621)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2617..2664
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2682..3497)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3405..3424
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3489..3508
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg atcccgcgaa attaatacga ctcactatag ggagaccaca
      661 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgcttaa
      721 acgttgtctc tctccgctca ccctggttaa tcaggttgcg cttattgtgt tgctttctac
      781 tgctattgga ctggcaggga tggcggtttc tggctggctg gtgcaaggcg ttcagggcag
      841 cgcccatgcg atcaacaaag cgggatcgct gcgcatgcaa agttaccgtc tgttggcggc
      901 agtgccatta agcgagaaag acaagccctt aattaaagag atggaacaaa cggcatttag
      961 cgccgagttg actcgagcag cagaacgaga cggacaactg gcgcaattac agggtttaca
     1021 agattactgg cgtaatgaac tgatccctgc gctgatgcgt gcacaaaacc gagaaacggt
     1081 gtcagcggat gtcagccagt ttgttgccgg gcttgatcaa ctggtatctg gttttgaccg
     1141 caccacggaa atgcgcatcg agacagtggt actggtccat cgggtaatgg cggtatttat
     1201 ggcactttta ctggtgttca ctattatctg gttgcgggcg cgactgctac aaccgtggcg
     1261 gcaactgctg gcaatggcga gtgccgtcag tcatcgcgat tttacccaac gcgcaaacat
     1321 cagcgggcgc aacgaaatgg cgatgcttgg aactgcgttg aacaatatgt ctgcagaact
     1381 ggccgaaagt tatgccgtac ttgagcagcg ggttcaggag aaaaccgccg ggctggagca
     1441 taaaaatcag atcctctctt ttttatggca ggctaaccgc cgtttgcatt cccgcgcccc
     1501 gctgtgtgaa cgcctgtcac ctgtactcaa cggcttacag aatttaaccc tgctacgtga
     1561 tatcgaattg cgggtgtatg acactgatga tgaagagaat catcaggagt ttacctgcca
     1621 gccagatatg acttgtgatg ataaaggctg ccagctctgc ccgcgcggcg tattacccgt
     1681 tggtgatcgc ggcacgaccc tgaagtggcg gctggctgac tctcatacgc agtacggtat
     1741 tttgctggcg accctgccac aggggcgtca tcttagccat gatcaacaac aactggtgga
     1801 taccctggct gaacaactca ccgccacgct ggcgctggat cgccatcagg aacgtcagca
     1861 acagttgatc gtgatggaag agcgtgccac cattgcgcgc gaactgcatg attctattgc
     1921 ccaatctctc tcttgcatga agatgcaggt gagttgttta cagatgcagg gcgatgcgct
     1981 gccagaaagc agccgcgaac tgttaagtca gatccgtaac gaactgaatg catcctgggc
     2041 gcagttgcgt gaattgctca tcacattccg cttgcagctc accgagcctg gattacgtcc
     2101 ggcgctggag gcgagttgcg aagagtacag cgccaaattt ggcttcccgg tgaagctgga
     2161 ttatcaattg ccgcctcgcc tggtgccttc gcatcaggca atccacttgt tgcaaattgc
     2221 ccgtgaggca ttaagtaacg ccctcaaaca ttcgcaagcg agtgaagtcg tggtgacggt
     2281 ggcgcaaaac gataatcagg tcaaactgac cgtccaggat aacggctgcg gcgtgcctga
     2341 aaatgccatc cgcagcaatc actacggcat gataataatg cgcgatcgtg cgcaaagttt
     2401 acgaggcgat tgccgcgtcc gccgtcgtga atcaggtggc accgaagtgg tggtcacctt
     2461 tattcccgaa aaaactttca cagacgtcca aggagatacc catgagggag gaggaagcga
     2521 gcagaaactc atctctgaag aggatctgta agtcgaccgg ctgctaacaa agcccgaaag
     2581 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2641 aaacgggtct tgaggggttt tttgctgaaa gccaattctg attagaaaaa ctcatcgagc
     2701 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2761 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2821 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2881 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2941 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     3001 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     3061 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     3121 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3181 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3241 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3301 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3361 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3421 tacccatata aatcagcatc catgttggaa tttaatcgcg gcttcgagca agacgtttcc
     3481 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     3541 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     3601 tg
//
