LOCUS       sequence-454026-        3734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a NarX-based chimera capable of sensing nickel (II). 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 3734)
  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 3734)
  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 NrsS NarX Chimera
FEATURES             Location/Qualifiers
     source          1..3734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..25
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6..24
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             56..78
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2024..2053
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2108..2126)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2122..2169
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2187..3002)
                     /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     2910..2929
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2994..3013
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3124..3712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3613..3632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt
       61 ttaactttaa gaaggagata tacatatgaa tacccgtcgt ctgtttgcac gtagccgtct
      121 gcagctggca ttttggtatg cactggttat gggtggtatt ctgaccctgt taggtctggg
      181 tgtttatcgt gcaattgttc aggcaaattg gatggcactg gaacgtgaag ttgaaagcat
      241 tgcaggcacc ctgcatgata gcctggaacc gatgctgccg agcaatgcaa gcccgaccgg
      301 tgtgctgcag aaaatgctgc cggatctgtg tctggttaat cagccgtgcc aggttaatcc
      361 gacactgatt gaacgtcata ccctgggtat tagcgatcgt agcctgtatt atatccgcct
      421 gtttgattat cagggtaatc tgctggcgtt tagcccgaat cagcctgcaa gcctgagcag
      481 catctttaat caagaaacct ggcagaccat tcatccgcct accggtgatc gttatcgtca
      541 gtttaccacc attctgcata gcgcaggtaa taccgataaa agcagctggg gttatctgca
      601 aattggtcgt agtctggcag cctttgatgc cgaaaataaa cgtattctgt ggattctggg
      661 tctgagcttt ccgattgcac tgggtttagt tgcatttagc agttggggtt tagcaggtct
      721 ggcaatgcgt ccgatttatc agagctatca gcagcagcaa cagtttaccg caaatgcagc
      781 acatgaactg cgtagtccgc tggcaagcct gctggcaacc gttgaagcag ttctgcgtat
      841 tgatagcagc catagtccgg aaattaacac catgtctgca gaactggccg aaagttatgc
      901 cgtacttgag cagcgggttc aggagaaaac cgccgggctg gagcataaaa atcagatcct
      961 ctctttttta tggcaggcta accgccgttt gcattcccgc gccccgctgt gtgaacgcct
     1021 gtcacctgta ctcaacggct tacagaattt aaccctgcta cgtgatatcg aattgcgggt
     1081 gtatgacact gatgatgaag agaatcatca ggagtttacc tgccagccag atatgacttg
     1141 tgatgataaa ggctgccagc tctgcccgcg cggcgtatta cccgttggtg atcgcggcac
     1201 gaccctgaag tggcggctgg ctgactctca tacgcagtac ggtattttgc tggcgaccct
     1261 gccacagggg cgtcatctta gccatgatca acaacaactg gtggataccc tggctgaaca
     1321 actcaccgcc acgctggcgc tggatcgcca tcaggaacgt cagcaacagt tgatcgtgat
     1381 ggaagagcgt gccaccattg cgcgcgaact gcatgattct attgcccaat ctctctcttg
     1441 catgaagatg caggtgagtt gtttacagat gcagggcgat gcgctgccag aaagcagccg
     1501 cgaactgtta agtcagatcc gtaacgaact gaatgcatcc tgggcgcagt tgcgtgaatt
     1561 gctcatcaca ttccgcttgc agctcaccga gcctggatta cgtccggcgc tggaggcgag
     1621 ttgcgaagag tacagcgcca aatttggctt cccggtgaag ctggattatc aattgccgcc
     1681 tcgcctggtg ccttcgcatc aggcaatcca cttgttgcaa attgcccgtg aggcattaag
     1741 taacgccctc aaacattcgc aagcgagtga agtcgtggtg acggtggcgc aaaacgataa
     1801 tcaggtcaaa ctgaccgtcc aggataacgg ctgcggcgtg cctgaaaatg ccatccgcag
     1861 caatcactac ggcatgataa taatgcgcga tcgtgcgcaa agtttacgag gcgattgccg
     1921 cgtccgccgt cgtgaatcag gtggcaccga agtggtggtc acctttattc ccgaaaaaac
     1981 tttcacagac gtccaaggag atacccatga gggaggagga agcgagcaga aactcatctc
     2041 tgaagaggat ctgtaagtcg accggctgct aacaaagccc gaaaggaagc tgagttggct
     2101 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     2161 ggttttttgc tgaaagccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc
     2221 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     2281 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     2341 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     2401 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt
     2461 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     2521 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     2581 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     2641 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     2701 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     2761 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     2821 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     2881 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     2941 gcatccatgt tggaatttaa tcgcggcttc gagcaagacg tttcccgttg aatatggctc
     3001 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     3061 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtgaga tcaaaggatc
     3121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3241 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3301 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3361 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3421 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3481 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3541 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3601 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3661 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3721 caacgcgatc ccgc
//
