LOCUS       sequence-455039-        3707 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses a NarX-based chimera capable of sensing copper (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 3707)
  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 3707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 CusS NarX Chimera
FEATURES             Location/Qualifiers
     source          1..3707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..24
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5..23
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             55..77
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1987..2016
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2071..2089)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2085..2132
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2150..2965)
                     /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     2873..2892
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2957..2976
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3096..3684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3585..3604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
       61 taactttaag aaggagatat acatatggtg agcaaaccgt ttcagcgtcc gtttagcctg
      121 gcaacccgtc tgaccttttt catttcactg gcaaccattg cagcattttt cgcatttgca
      181 tggattatga tccatagcgt gaaagtgcat tttgccgaac aggatattaa cgacctgaaa
      241 gaaattagcg caaccctgga acgtgttctg aatcatcctg atgaaaccca ggcacgtcgt
      301 ctgatgaccc tggaagatat tgttagcggt tatagcaatg ttctgattag tctggcagat
      361 agccagggta aaaccgttta tcatagtccg ggtgcaccgg atattcgtga atttacccgt
      421 gatgcaattc cggataaaga tgcacaaggt ggtgaagttt atctgctgag cggtccgacc
      481 atgatgatgc ctggtcatgg tcatggccac atggaacata gcaattggcg tatgattaat
      541 ctgccggttg gtccgctggt tgatggtaaa ccgatttata ccctgtatat tgccctgagc
      601 attgattttc atctgcacta tatcaacgat ctgatgaaca aactgattat gaccgcaagc
      661 gttattagca tcctgattgt ttttattgtt ctgctggcag ttcataaagg tcatgcaccg
      721 attcgtagcg ttagccgtca gattcagaat attaccagca aagatctgga tgttcgtctg
      781 gatccgcaga ccgttccgat tgaactggaa cagctggttc tgagctttaa tcatatgagc
      841 gcagaactgg cagaaagcta tgcagttctg gaacagcgtg ttcaagagaa aaccgcaggc
      901 ctggaacata aaaatcagat tctgagcttt ctgtggcagg caaatcgtcg tctgcatagc
      961 cgtgcaccgc tgtgtgaacg tctgagtccg gttctgaatg gtctgcagaa cctgacactg
     1021 ctgcgtgata ttgaactgcg tgtttatgat accgatgatg aagaaaacca ccaagaattt
     1081 acctgtcagc cggatatgac ctgtgatgat aaaggttgtc agctgtgtcc gcgtggtgtt
     1141 ctgccggtgg gtgatcgtgg tacaaccctg aaatggcgtc tggccgatag tcatacccag
     1201 tatggtatcc tgctggcgac actgccgcag ggtcgtcatc tgagccatga tcagcagcag
     1261 ctggtagata ccctggccga acagctgacc gcaacactgg cactggatcg tcatcaagaa
     1321 cgtcagcagc aactgattgt tatggaagaa cgtgccacca ttgcgcgtga actgcatgat
     1381 agcattgcac agagcctgag ctgtatgaaa atgcaggtta gctgtctgca gatgcagggt
     1441 gatgcactgc cggaaagctc acgcgaactg ctgagccaga ttcgcaatga actgaatgcc
     1501 agctgggcac agctgcgtga gctgctgatt acctttcgtc tgcagctgac agaaccgggt
     1561 ctgcgtccgg cactggaagc aagctgtgaa gaatattcag cgaaatttgg ttttccggtg
     1621 aaactggatt atcagctgcc tccgcgtctg gttccgagcc atcaggcaat tcatctgctg
     1681 caaattgcac gtgaagcact gagcaatgca ctgaaacatt cacaggcaag cgaagttgtt
     1741 gttaccgttg cacagaatga taaccaggtt aaactgaccg ttcaggataa tggttgtggt
     1801 gttccggaaa atgcaattcg tagcaatcat tatggcatga tcattatgcg tgatcgtgcc
     1861 cagagcctgc gtggtgattg tcgtgttcgt cgtcgtgaaa gcggtggtac agaagtggtg
     1921 gttaccttta ttcctgagaa aacctttacc gatgttcagg gtgataccca tgaaggtggt
     1981 ggtagcgaac agaaactgat ttcagaagaa gatctgtaag tcgaccggct gctaacaaag
     2041 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     2101 gggcctctaa acgggtcttg aggggttttt tgctgaaagc caattctgat tagaaaaact
     2161 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     2221 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2281 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2341 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2401 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2461 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2521 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2581 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2641 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2701 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2761 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2821 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2881 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcgagcaag
     2941 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     3001 gttttattgt tcatgatgat atatttttat cttgtgcaac ttgtgcaatg taacatcaga
     3061 gattttgaga cacaacgtga gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3301 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3361 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3421 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3481 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3541 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3601 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3661 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcga tcccgcg
//
