LOCUS       sequence-241980-        4269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An arsenic sensor with optimized intracellular receptor density. 
            Also used in the sensor cell array. pSB3K3 carrying 
            J117-30arsR-t-ParsR-30gfp-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4269)
  AUTHORS   Wan X, Volpetti F, Petrova E, French C, Maerkl SJ, Wang B
  TITLE     Cascaded amplifying circuits enable ultrasensitive cellular sensors 
            for toxic metals.
  JOURNAL   Nat Chem Biol. 2019 May;15(5):540-548. doi: 
            10.1038/s41589-019-0244-3. Epub 2019 Mar 25.
  PUBMED    30911179
REFERENCE   2  (bases 1 to 4269)
  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"; doi: 
            "10.1038/s41589-019-0244-3"; journalName: "Nat Chem Biol"; date: 
            "2019-05"; volume: "15"; issue: "5"; pages: "540-548"
COMMENT     Sequence Label: As3 (pBW411J117-arsR)
FEATURES             Location/Qualifiers
     source          1..4269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(141..956)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPYSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     864..883
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     948..967
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(1551..2095)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1630..1649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2563..2581
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2585..2628)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2631..2652
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        2653..2687
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             2699..2710
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3133..3204
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3220..3247
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3375..3386
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3393..4106
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3423..3451)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4046..4066
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      4129..4200
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4216..4243
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    join(4250..4269,1)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
       61 tcactcaaag gcggtaatct cgagtcccgt caagtcagcg taatgctctg ccagtgttac
      121 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
      181 ttcatatcag gattatcgat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
      241 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
      301 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
      361 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
      421 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
      481 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
      541 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
      601 ttttcacctg aataaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
      661 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
      721 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
      781 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
      841 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
      901 atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca
      961 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     1021 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     1081 acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg
     1141 gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt
     1201 ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca
     1261 ccttcttcac gaggcagacc tcagcgctag cggagtgtat actggcttac tatgttggca
     1321 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     1381 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     1441 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     1501 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     1561 ctccgccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     1621 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     1681 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     1741 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     1801 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     1861 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     1921 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     1981 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcaaaaaac cgccctgcaa
     2041 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     2101 tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattccat ggtgccacct
     2521 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2581 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg
     2641 ccgcttctag agttgacagc tagctcagtc ctagggattg tgctagctac tagagattaa
     2701 agaggagaaa tactagatgt catttctgtt acccatccaa ttgttcaaaa ttcttgctga
     2761 tgaaacccgt ctgggcatcg ttttactgct cagcgaactg ggagagttat gcgtctgcga
     2821 tctctgcact gctctcgacc agtcgcagcc caagatctcc cgccacctgg cattgctgcg
     2881 tgaaagcggg ctattgctgg accgcaagca aggtaagtgg gttcattacc gcttatcacc
     2941 gcatattcca gcatgggcgg cgaaaattat tgatgaggcc tggcgatgtg aacaggaaaa
     3001 ggttcaggcg attgtccgca acctggctcg acaaaactgt tccggggaca gtaagaacat
     3061 ttgcagttaa aaatttagct aaacacacat gaattttcag atgtgtttta tccgggtact
     3121 agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     3181 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     3241 ttctgcgttt atatactaga gccaactcaa aattcacacc tattaccttc ctctgcactt
     3301 acacattcgt taagtcatat atgtttttga cttatccgct tcgaagagag acactacctg
     3361 caatactaga gattaaagag gagaaatact agatgcgtaa aggagaagaa cttttcactg
     3421 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     3481 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     3541 ctggaaaact acctgttcca tggccaacac ttgtcactac tttcggttat ggtgttcaat
     3601 gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg
     3661 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     3721 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     3781 ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac tcacacaatg
     3841 tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca
     3901 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     3961 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     4021 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     4081 cacatggcat ggatgaacta tacaaataat aatactagag ccaggcatca aataaaacga
     4141 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     4201 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat actagtagcg
     4261 gccgctgca
//
