LOCUS       sequence-442138-        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"
     RBS             2..24
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1592..1621
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1676..1694)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1690..1737
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1755..2570)
                     /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     2478..2497
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2562..2581
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2692..3280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3181..3200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3308..3327
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3308..3326
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttttgtttaa ctttaagaag gagatataca tatggtgatc aagctgaaga acaagaaaaa
       61 cgactacagc aaactggaac gcaagctgta tatgtatatt gttgccattg ttgtggtggc
      121 cattgtgttt gttctgtata ttcgtagcat gatccgtggc aaattaggtg attggattct
      181 gagcattctg gaaaacaaat atgatctgaa tcacctggat gccatgaaac tgtatcagta
      241 tagcattcgc aacaacatcg acatctttat ctatgtggcg attgtgatta gcattctgat
      301 tctgtgtcgt gtgatgctga gtaaattcgc caaatatttc gatgaaatca acaccggcat
      361 tgatgtgctg attcagaatg aagataagca gattgaactg agcgcagaaa tggatgttat
      421 ggaacagaaa ctgaacacca tgtctgcaga actggccgaa agttatgccg tacttgagca
      481 gcgggttcag gagaaaaccg ccgggctgga gcataaaaat cagatcctct cttttttatg
      541 gcaggctaac cgccgtttgc attcccgcgc cccgctgtgt gaacgcctgt cacctgtact
      601 caacggctta cagaatttaa ccctgctacg tgatatcgaa ttgcgggtgt atgacactga
      661 tgatgaagag aatcatcagg agtttacctg ccagccagat atgacttgtg atgataaagg
      721 ctgccagctc tgcccgcgcg gcgtattacc cgttggtgat cgcggcacga ccctgaagtg
      781 gcggctggct gactctcata cgcagtacgg tattttgctg gcgaccctgc cacaggggcg
      841 tcatcttagc catgatcaac aacaactggt ggataccctg gctgaacaac tcaccgccac
      901 gctggcgctg gatcgccatc aggaacgtca gcaacagttg atcgtgatgg aagagcgtgc
      961 caccattgcg cgcgaactgc atgattctat tgcccaatct ctctcttgca tgaagatgca
     1021 ggtgagttgt ttacagatgc agggcgatgc gctgccagaa agcagccgcg aactgttaag
     1081 tcagatccgt aacgaactga atgcatcctg ggcgcagttg cgtgaattgc tcatcacatt
     1141 ccgcttgcag ctcaccgagc ctggattacg tccggcgctg gaggcgagtt gcgaagagta
     1201 cagcgccaaa tttggcttcc cggtgaagct ggattatcaa ttgccgcctc gcctggtgcc
     1261 ttcgcatcag gcaatccact tgttgcaaat tgcccgtgag gcattaagta acgccctcaa
     1321 acattcgcaa gcgagtgaag tcgtggtgac ggtggcgcaa aacgataatc aggtcaaact
     1381 gaccgtccag gataacggct gcggcgtgcc tgaaaatgcc atccgcagca atcactacgg
     1441 catgataata atgcgcgatc gtgcgcaaag tttacgaggc gattgccgcg tccgccgtcg
     1501 tgaatcaggt ggcaccgaag tggtggtcac ctttattccc gaaaaaactt tcacagacgt
     1561 ccaaggagat acccatgagg gaggaggaag cgagcagaaa ctcatctctg aagaggatct
     1621 gtaagtcgac cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1681 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     1741 aaagccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     1801 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     1861 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     1921 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     1981 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     2041 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     2101 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     2161 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     2221 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     2281 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     2341 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     2401 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     2461 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     2521 gaatttaatc gcggcttcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     2581 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     2641 gcaatgtaac atcagagatt ttgagacaca acgtgagatc aaaggatctt cttgagatcc
     2701 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2761 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2821 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2881 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2941 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3001 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3061 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3121 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3181 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3241 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcgatccc
     3301 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag aaataa
//
