LOCUS       sequence-243982-        8036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arsenic sensor used in the sensor cell array. pSB4A3 carrying 
            J109-30arsR-t-ParsR-ABS67-30hrpR-30hrpS-t-PhrpLE-30RinA-t-PrinA-30gf
            p-t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8036)
  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 8036)
  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: As0
FEATURES             Location/Qualifiers
     source          1..8036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2..59
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(83..100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(362..1312)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1360..1582
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2074..2168)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2198..2989,2990..3058))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2821..2840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3059..3163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3228..3246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3250..3293)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3296..3317
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        3318..3352
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     RBS             3364..3375
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3798..3869
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3885..3912
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             4086..4097
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5008..5019
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             5082..5093
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6028..6099
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6115..6142
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             6376..6387
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6833..6904
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6920..6947
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             7142..7153
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7160..7873
                     /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(7190..7218)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7813..7833
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      7896..7967
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7983..8010
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    join(8017..8036,1)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
       61 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatctcgag gttacattgt cgatctgttc
      181 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
      241 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
      301 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
      361 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
      421 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
      481 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
      541 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
      601 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
      661 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
      721 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
      781 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
      841 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
      901 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
      961 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     1021 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     1081 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
     1141 tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
     1201 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
     1261 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
     1321 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     1381 atgtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac
     1441 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     1501 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     1561 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     1621 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     1681 ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg
     1741 agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg
     1801 ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa
     1861 gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc
     1921 tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt
     1981 atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac
     2041 aggcttaccc gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg
     2101 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     2161 caggagtcca agcgagctcg taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt cccatggtgc
     3181 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3241 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt
     3301 cgcggccgct tctagagttt acagctagct cagtcctagg gactgtgcta gctactagag
     3361 attaaagagg agaaatacta gatgtcattt ctgttaccca tccaattgtt caaaattctt
     3421 gctgatgaaa cccgtctggg catcgtttta ctgctcagcg aactgggaga gttatgcgtc
     3481 tgcgatctct gcactgctct cgaccagtcg cagcccaaga tctcccgcca cctggcattg
     3541 ctgcgtgaaa gcgggctatt gctggaccgc aagcaaggta agtgggttca ttaccgctta
     3601 tcaccgcata ttccagcatg ggcggcgaaa attattgatg aggcctggcg atgtgaacag
     3661 gaaaaggttc aggcgattgt ccgcaacctg gctcgacaaa actgttccgg ggacagtaag
     3721 aacatttgca gttaaaaatt tagctaaaca cacatgaatt ttcagatgtg ttttatccgg
     3781 gtactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     3841 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     3901 ggcctttctg cgtttatata ctagagccaa ctcaaaattc acacctatta ccttcctctg
     3961 cacttacaca ttcgttaagt catatatgtt tttgacttat ccgcttcgaa gagagacact
     4021 acctgcaaac ttacacattc gttaagtcat atatgttttt gacttatccg cttctactag
     4081 agattaaaga ggagaaatac tagatgagta caggcatcga taaggacgtc cgagagtgtt
     4141 ggggcgtaac tgcattatca gcgggtcatc aaattgcaat gaatagcgcg tttctggata
     4201 tggacttgct gttgtgcggg gaaaccggca ccggcaagga cacactggcc aaccgcattc
     4261 acgagttgtc cagcaggtcg ggaccctttg tgggcatgaa ctgcgccgcc attcccgagt
     4321 cgctggcaga gagccagtta ttcggtgtgg tcaacggtgc attcaccggc gtatgccggg
     4381 ctcgcgaggg ctacatagag gcctccagtg gtggcacctt gtacctggat gaaatcgaca
     4441 gcatgccgtt gagcctgcaa gccaaactgc tgcgtgtgtt ggagagtcga ggtatcgagc
     4501 gtctgggctc gaccgaattt atcccggtgg atctgcggat cattgcctcg gcccagcggc
     4561 cactggatga actggtggaa caaggacttt tccgtcgcga cctgtttttt cggctcaacg
     4621 tgctgacgct tcacttgcca gccttgcgca aacgtcgtga acagatcctg ccattgttcg
     4681 accagttcac ccagggtatc gctgccgagt tcggacgtcc cgctcctgcg ctggacagcg
     4741 ggcgtgtgca gctgctgctc agccacgact ggccgggcaa catccgcgaa ttgaagtctg
     4801 cggccaagcg cttcgtactc ggcttcccct tgctgggcgc cgaccctgtg gaagcgcttg
     4861 accctgccac ggggctgcgc acgcaaatgc gcatcatcga gaaaatgctc atccaggatg
     4921 ccttgaagcg gcacaggcac aatttcgacg cggtgcttca ggagttggag ttgccaagac
     4981 gcaccctgta tcaccgcatg aaggaactgg gagttgcagc gccgatcgct gcgacggccg
     5041 gggtctaata aaacgcaggt gtggttatgc tactagagat taaagaggag aaatactaga
     5101 tgagtcttga tgaaaggttt gaggatgatc tggacgagga gcgggttccg aatctgggga
     5161 tagttgccga aagtatttcg caactgggta tcgacgtgct gctatcgggt gagaccggca
     5221 cgggcaaaga cacgattgcc cgacggattc atgagatgtc aggccgcaaa gggcgcctgg
     5281 tggcgatgaa ttgcgcggcc attccggagt ccctcgccga gagcgagtta ttcggcgtgg
     5341 tcagcggtgc ctacaccggc gctgatcgct ccagagtcgg ttatgtcgaa gcggcgcagg
     5401 gcggcacgct gtacctggat gagatcgata gcatgccgct gagcctgcaa gccaaattgc
     5461 tgagggtgct ggaaacccga gcgcttgaac ggctgggttc gacgtcgacg atcaagctgg
     5521 atatctgcgt gatcgcctcc gcccaatgct cgctggacga cgccgtcgag cgggggcagt
     5581 ttcgtcgcga tctgtatttt cgcctgaacg tcctgacact caagcttcct ccgctacgta
     5641 accagtctga tcgcatagtt cccctgttca cacgttttac ggccgccgcc gcgagggagc
     5701 tcggtgttcc cgttcccgat gtttgcccac tgctgcacaa agtgctgctg ggccacgact
     5761 ggcccggcaa tatccgtgag ctcaaggcgg cagccaaacg ccatgtgctg ggtttcccct
     5821 tgctgggcgc cgagccgcag ggcgaagagc acttggcctg tgggctcaaa tcgcaattgc
     5881 gagtgatcga aaaagccctg attcaggagt cgctcaagcg ccacgacaat tgtgtggatt
     5941 cggtaagcct ggaactggac gtgccacgcc gtacgctcta tcgacgcatc aaagaattgc
     6001 agatctaata atactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     6061 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     6121 ccttcgggtg ggcctttctg cgtttatata ctagaggccg gattatgtcc gctgagtggg
     6181 tcacggtccc ggatcagttc ccttgcgaag ctgaccgatg tttttgtgcc aaaagctgtt
     6241 gtggcaaaaa acggtttgcg caaagttttg tattacaaag aatttcacat tttaaaatat
     6301 ctttataaat caatcagtta tttctatttt caagctggca cggttattgc tatagggctt
     6361 gtactactag agattaaaga ggagaaatac tagatgacta aaaagaaata tggattaaaa
     6421 ttatcaacag ttcgaaagtt agaagatgag ttgtgtgatt atcctaatta tcataagcaa
     6481 ctcgaagatt taagaagtga aataatgaca ccatggattc caacagatac aaatataggc
     6541 ggggagtttg taccgtctaa tacatcgaaa acagaaatgg cagtaactaa ttatctttgt
     6601 agtatacgaa gaggtaaaat ccttgagttt aagagcgcta ttgaacgtat aatcaacaca
     6661 tcaagtagga aagaacgcga atttattcaa gagtattatt ttaataaaaa ggaattagtg
     6721 aaagtttgtg atgacataca catttctgat agaactgctc atagaatcaa aaggaaaatc
     6781 atatccagat tggcggaaga gttaggggaa gagtgatact agagccaggc atcaaataaa
     6841 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     6901 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactaga
     6961 gaattggcag taaagtggca gtttttgata cctaaaatga gatattatga tagtgtagga
     7021 tattgactat cttactgcgt ttcccttatc gcaattagga ataaaggatc tatgtgggtt
     7081 ggctgattat agccaatcct tttttaattt taaaaagcgt atagcgcgag tactagagat
     7141 taaagaggag aaatactaga tgcgtaaagg agaagaactt ttcactggag ttgtcccaat
     7201 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     7261 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     7321 tgttccatgg ccaacacttg tcactacttt cggttatggt gttcaatgct ttgcgagata
     7381 cccagatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     7441 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
     7501 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     7561 aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     7621 agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
     7681 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     7741 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     7801 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     7861 tgaactatac aaataataat actagagcca ggcatcaaat aaaacgaaag gctcagtcga
     7921 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     7981 ctggctcacc ttcgggtggg cctttctgcg tttatatact agtagcggcc gctgca
//
