LOCUS       sequence-454023-        3356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a NarX-based chimera capable of sensing vancomycin. 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 3356)
  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 3356)
  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 VanS NarX Chimera
FEATURES             Location/Qualifiers
     source          1..3356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..35
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        16..34
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             66..88
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1656..1685
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1740..1758)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1754..1801
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1819..2634)
                     /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     2542..2561
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2626..2645
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2756..3344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3245..3264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa
       61 ataattttgt ttaactttaa gaaggagata tacatatggt gatcaagctg aagaacaaga
      121 aaaacgacta cagcaaactg gaacgcaagc tgtatatgta tattgttgcc attgttgtgg
      181 tggccattgt gtttgttctg tatattcgta gcatgatccg tggcaaatta ggtgattgga
      241 ttctgagcat tctggaaaac aaatatgatc tgaatcacct ggatgccatg aaactgtatc
      301 agtatagcat tcgcaacaac atcgacatct ttatctatgt ggcgattgtg attagcattc
      361 tgattctgtg tcgtgtgatg ctgagtaaat tcgccaaata tttcgatgaa atcaacaccg
      421 gcattgatgt gctgattcag aatgaagata agcagattga actgagcgca gaaatggatg
      481 ttatggaaca gaaactgaac accatgtctg cagaactggc cgaaagttat gccgtacttg
      541 agcagcgggt tcaggagaaa accgccgggc tggagcataa aaatcagatc ctctcttttt
      601 tatggcaggc taaccgccgt ttgcattccc gcgccccgct gtgtgaacgc ctgtcacctg
      661 tactcaacgg cttacagaat ttaaccctgc tacgtgatat cgaattgcgg gtgtatgaca
      721 ctgatgatga agagaatcat caggagttta cctgccagcc agatatgact tgtgatgata
      781 aaggctgcca gctctgcccg cgcggcgtat tacccgttgg tgatcgcggc acgaccctga
      841 agtggcggct ggctgactct catacgcagt acggtatttt gctggcgacc ctgccacagg
      901 ggcgtcatct tagccatgat caacaacaac tggtggatac cctggctgaa caactcaccg
      961 ccacgctggc gctggatcgc catcaggaac gtcagcaaca gttgatcgtg atggaagagc
     1021 gtgccaccat tgcgcgcgaa ctgcatgatt ctattgccca atctctctct tgcatgaaga
     1081 tgcaggtgag ttgtttacag atgcagggcg atgcgctgcc agaaagcagc cgcgaactgt
     1141 taagtcagat ccgtaacgaa ctgaatgcat cctgggcgca gttgcgtgaa ttgctcatca
     1201 cattccgctt gcagctcacc gagcctggat tacgtccggc gctggaggcg agttgcgaag
     1261 agtacagcgc caaatttggc ttcccggtga agctggatta tcaattgccg cctcgcctgg
     1321 tgccttcgca tcaggcaatc cacttgttgc aaattgcccg tgaggcatta agtaacgccc
     1381 tcaaacattc gcaagcgagt gaagtcgtgg tgacggtggc gcaaaacgat aatcaggtca
     1441 aactgaccgt ccaggataac ggctgcggcg tgcctgaaaa tgccatccgc agcaatcact
     1501 acggcatgat aataatgcgc gatcgtgcgc aaagtttacg aggcgattgc cgcgtccgcc
     1561 gtcgtgaatc aggtggcacc gaagtggtgg tcacctttat tcccgaaaaa actttcacag
     1621 acgtccaagg agatacccat gagggaggag gaagcgagca gaaactcatc tctgaagagg
     1681 atctgtaagt cgaccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1741 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1801 gctgaaagcc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     1861 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1921 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1981 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2041 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     2101 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2161 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     2221 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     2281 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     2341 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     2401 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     2461 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     2521 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     2581 gttggaattt aatcgcggct tcgagcaaga cgtttcccgt tgaatatggc tcataacacc
     2641 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     2701 ttgtgcaatg taacatcaga gattttgaga cacaacgtga gatcaaagga tcttcttgag
     2761 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2821 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2881 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2941 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3001 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3061 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3121 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3181 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3241 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3301 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaac
//
