LOCUS       sequence-454890-        2459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NarL with a mutated DNA binding domain (YdfI). For use 
            with NarX chimeras and reporter plasmids from this study..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2459)
  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 2459)
  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 NarL YdfI Flag
FEATURES             Location/Qualifiers
     source          1..2459
                     /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             1382..1405
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1460..1478)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1474..1521
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1539..2354)
                     /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     2262..2281
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2346..2365
                     /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 atatgagtaa
      721 tcaggaaccg gctactatcc tgctgattga cgatcacccg atgctgcgaa ctggcgtaaa
      781 acagcttatc agtatggcac cagatatcac cgtggttggc gaagcgagta atggcgaaca
      841 gggtattgaa ctggcggagt ctcttgatcc cgatctgatc ctgttagatc tcaatatgcc
      901 cggcatgaac ggtctggaaa cgctggataa actgcgcgaa aagtccctct cagggcgcat
      961 tgtggtattc agcgtctcta accatgaaga agatgtggtc accgcactga aacgcggcgc
     1021 ggatggctat ctgttaaaag atatggaacc ggaagatctg ctgaaagcat tgcatcaggc
     1081 agctgctggc gaaatggtat taagccctga tatccttaaa cgtctgcaag aaatccaatt
     1141 tgagcggatg aaaaagcagc gcaatgagac gcagctgaca gaaaaggaag tcattgttct
     1201 aaaagcaatt gctaaaggtc ttaaaagcaa agcgattgcc tttgatttgg gcgtctctga
     1261 gcgaacagta aagtccagat taacgtccat ttacaataaa ttaggcgcga attcaagaac
     1321 tgaggcagta acgattgcca tgcaaaaagg tattctgaca atagacaacg gaggaggaag
     1381 cgattacaag gatgacgacg ataagtaagt cgaccggctg ctaacaaagc ccgaaaggaa
     1441 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1501 cgggtcttga ggggtttttt gctgaaagcc aattctgatt agaaaaactc atcgagcatc
     1561 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     1621 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     1681 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     1741 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     1801 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     1861 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     1921 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     1981 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     2041 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     2101 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     2161 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     2221 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     2281 ccatataaat cagcatccat gttggaattt aatcgcggct tcgagcaaga cgtttcccgt
     2341 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     2401 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtg
//
