LOCUS       sequence-454025-        3629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a NarX-based chimera capable of sensing iron (III). 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 3629)
  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 3629)
  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 RssA NarX Chimera
FEATURES             Location/Qualifiers
     source          1..3629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..23
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4..22
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             54..76
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1917..1946
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2001..2019)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2015..2062
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2080..2895)
                     /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     2803..2822
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2887..2906
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3017..3605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3506..3525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt
       61 aactttaaga aggagatata catatgatcg gcttcaaaag cttctttatg cgcaccatta
      121 tctttcaggt tctggcaatt ctgctgctgt ggggtctgct ggttgcatgg gtgaaatatt
      181 ggtattatcc ggacatggaa aaatactttg ataaccagca gcgtattgtt gcagcaggta
      241 ttgcaaatat tctggatgaa accggcaccg acaatattga ttatcgcggt attatcaaaa
      301 ccatcgaggg catgtatatc gacagcatta ataacggtat gcaggatgag atcgattatc
      361 atccgctgtt tgttgtgtat gatcgtgata atcgtgtgct gtatagcagt cagacccagg
      421 gtgaaccgct gcgtctgcct ccgagcgttc tgagcggtag cgttaattat gccggtgcaa
      481 attggcatct ggcaggtagc tggaaagaaa aacgtcagta tcgtgttatt gtgggcgaaa
      541 gctttaatga tcgtaccaca ctgtttggta atccggcaga tgttccgctg ctgggtattc
      601 tggcagcaat tattgttacc ctgctgttta ccgcatattt cagcctgcgt ccgctgcgcc
      661 agattgcacg taccattagc gatcgtcagc ctggtaatct gagcccgatt aatgttagcg
      721 aacagtatca agaaattcgt ccggttgtta tggaagtgaa caaaatgtct gcagaactgg
      781 ccgaaagtta tgccgtactt gagcagcggg ttcaggagaa aaccgccggg ctggagcata
      841 aaaatcagat cctctctttt ttatggcagg ctaaccgccg tttgcattcc cgcgccccgc
      901 tgtgtgaacg cctgtcacct gtactcaacg gcttacagaa tttaaccctg ctacgtgata
      961 tcgaattgcg ggtgtatgac actgatgatg aagagaatca tcaggagttt acctgccagc
     1021 cagatatgac ttgtgatgat aaaggctgcc agctctgccc gcgcggcgta ttacccgttg
     1081 gtgatcgcgg cacgaccctg aagtggcggc tggctgactc tcatacgcag tacggtattt
     1141 tgctggcgac cctgccacag gggcgtcatc ttagccatga tcaacaacaa ctggtggata
     1201 ccctggctga acaactcacc gccacgctgg cgctggatcg ccatcaggaa cgtcagcaac
     1261 agttgatcgt gatggaagag cgtgccacca ttgcgcgcga actgcatgat tctattgccc
     1321 aatctctctc ttgcatgaag atgcaggtga gttgtttaca gatgcagggc gatgcgctgc
     1381 cagaaagcag ccgcgaactg ttaagtcaga tccgtaacga actgaatgca tcctgggcgc
     1441 agttgcgtga attgctcatc acattccgct tgcagctcac cgagcctgga ttacgtccgg
     1501 cgctggaggc gagttgcgaa gagtacagcg ccaaatttgg cttcccggtg aagctggatt
     1561 atcaattgcc gcctcgcctg gtgccttcgc atcaggcaat ccacttgttg caaattgccc
     1621 gtgaggcatt aagtaacgcc ctcaaacatt cgcaagcgag tgaagtcgtg gtgacggtgg
     1681 cgcaaaacga taatcaggtc aaactgaccg tccaggataa cggctgcggc gtgcctgaaa
     1741 atgccatccg cagcaatcac tacggcatga taataatgcg cgatcgtgcg caaagtttac
     1801 gaggcgattg ccgcgtccgc cgtcgtgaat caggtggcac cgaagtggtg gtcaccttta
     1861 ttcccgaaaa aactttcaca gacgtccaag gagataccca tgagggagga ggaagcgagc
     1921 agaaactcat ctctgaagag gatctgtaag tcgaccggct gctaacaaag cccgaaagga
     1981 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     2041 acgggtcttg aggggttttt tgctgaaagc caattctgat tagaaaaact catcgagcat
     2101 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     2161 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     2221 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     2281 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     2341 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     2401 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     2461 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     2521 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     2581 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     2641 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     2701 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     2761 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     2821 cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcgagcaag acgtttcccg
     2881 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     2941 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     3001 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3061 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3601 ggaaaaacgc cagcaacgcg atcccgcga
//
