LOCUS       sequence-241757-        4269 bp    DNA     circular SYN 15-APR-2025
DEFINITION  An arsenic sensor with optimized intracellular receptor density. 
            pSB3K3 carrying J109-30arsR-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 Apr 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: pXWJ109As-gfp
FEATURES             Location/Qualifiers
     source          1..4269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..56
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     RBS             68..79
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      502..573
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      589..616
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             744..755
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             762..1475
                     /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(792..820)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1415..1435
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1498..1569
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1585..1612
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1619..1639
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(1779..2594)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1779..2594)
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2502..2521
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2586..2605
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(3189..3733)
                     /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(3268..3287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4201..4219
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4223..4266)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(4269,1..21)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aattcgcggc cgcttctaga gtttacagct agctcagtcc tagggactgt gctagctact
       61 agagattaaa gaggagaaat actagatgtc atttctgtta cccatccaat tgttcaaaat
      121 tcttgctgat gaaacccgtc tgggcatcgt tttactgctc agcgaactgg gagagttatg
      181 cgtctgcgat ctctgcactg ctctcgacca gtcgcagccc aagatctccc gccacctggc
      241 attgctgcgt gaaagcgggc tattgctgga ccgcaagcaa ggtaagtggg ttcattaccg
      301 cttatcaccg catattccag catgggcggc gaaaattatt gatgaggcct ggcgatgtga
      361 acaggaaaag gttcaggcga ttgtccgcaa cctggctcga caaaactgtt ccggggacag
      421 taagaacatt tgcagttaaa aatttagcta aacacacatg aattttcaga tgtgttttat
      481 ccgggtacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      541 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
      601 ggtgggcctt tctgcgttta tatactagag ccaactcaaa attcacacct attaccttcc
      661 tctgcactta cacattcgtt aagtcatata tgtttttgac ttatccgctt cgaagagaga
      721 cactacctgc aatactagag attaaagagg agaaatacta gatgcgtaaa ggagaagaac
      781 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
      841 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
      901 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ttcggttatg
      961 gtgttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt ttcaagagtg
     1021 ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca
     1081 agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
     1141 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
     1201 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa
     1261 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     1321 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     1381 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     1441 ctgggattac acatggcatg gatgaactat acaaataata atactagagc caggcatcaa
     1501 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1561 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatata
     1621 ctagtagcgg ccgctgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1681 gcggcgagcg gtatcagctc actcaaaggc ggtaatctcg agtcccgtca agtcagcgta
     1741 atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
     1801 aaatgaaact gcaatttatt catatcagga ttatcgatac catatttttg aaaaagccgt
     1861 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     1921 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     1981 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     2041 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     2101 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     2161 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     2221 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     2281 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     2341 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     2401 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     2461 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     2521 ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt
     2581 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     2641 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
     2701 ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac
     2761 aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac
     2821 aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc
     2881 tggtatgagt cagcaacacc ttcttcacga ggcagacctc agcgctagcg gagtgtatac
     2941 tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa
     3001 aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct
     3061 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg
     3121 cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca
     3181 aagccgtttt tccataggct ccgccccctg acaagcatca cgaaatctga cgctcaaatc
     3241 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     3301 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     3361 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     3421 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     3481 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     3541 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     3601 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     3661 caaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3721 aaaacgatct caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc
     3781 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3841 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3901 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3961 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4021 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4081 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4141 tattccatgg tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     4201 aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat
     4261 gatttctgg
//
