LOCUS       sequence-318577-        6325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Two-layered mercury responsive circuit carrying an ECF11-based 
            amplifier.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6325)
  AUTHORS   Wan X, Pinto F, Yu L, Wang B
  TITLE     Synthetic protein-binding DNA sponge as a tool to tune gene 
            expression and mitigate protein toxicity.
  JOURNAL   Nat Commun. 2020 Nov 24;11(1):5961. doi: 10.1038/s41467-020-19552-9.
  PUBMED    33235249
REFERENCE   2  (bases 1 to 6325)
  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: "Nat 
            Commun."; date: "2020-11-24"; volume: "11(1):5961. doi"; pages: " 
            10.1038/s41467-020-19552-9"
COMMENT     Sequence Label: pXW109PmerT-Pecf11-gfp
FEATURES             Location/Qualifiers
     source          1..6325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    56..77
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        78..112
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     terminator      591..662
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      678..705
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1406..1477
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1493..1520
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1606..1617
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1624..2337
                     /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(1654..1682)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2277..2297
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2360..2431
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2447..2474
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2481..2501
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2556..2650
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        2723..2814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2815..2883,2884..3675)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3679..3718
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            3865..3973
                     /note="incP origin of transfer"
     CDS             complement(4034..5182)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      5575..6106
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(6233..6319)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgcggccgct tctagagttt acagctagct cagtcctagg gactgtgcta gctactagag
      121 tcacacagga aagtactaga tggaaaataa tttggaaaac ctgaccattg gcgtttttgc
      181 caaggcggcc ggggtcaacg tggagacaat ccgcttctat cagcgcaagg gcctgttgcg
      241 ggaaccggac aagccttacg gcagcatccg ccgctatggg gaggcggacg tggttcgggt
      301 gaaattcgtg aaatcggcac agcggctggg gttcagtctg gacgagattg ccgagctgtt
      361 gcggctcgac gatggcaccc actgcgagga ggccagcagc ctggccgaac acaagctcaa
      421 ggacgtgcgc gagaagatgg ccgacttggc gcgcatggaa accgtgctgt ctgaactcgt
      481 gtgcgcctgc catgcacgaa aggggaatgt ttcctgcccg ttgatcgcgt cactacaggg
      541 cgaagcaggc ctggcaaggt cagctatgcc ttagtactag agccaggcat caaataaaac
      601 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
      661 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atggatccgt
      721 tccatatcgc ttgactacgt acatgagtac ggaagtaagg ttacgctatc caatcctact
      781 agagtcacac aggactacta gatgatgagc gatagtccgc agaaactggg tcgtaatgaa
      841 tggaatgcat atatggataa agtgaaagcc aaagatcgtg aagcctttgc ctttgtgttt
      901 cgtttttatg caccgaaact gaaacagttc gcctataaac atgttggcaa tgaacaggtt
      961 gcaatggaaa tggttcaaga aaccatggca accgtttggc agaaagcaca tctgtatgat
     1021 ggtaaaaaaa gcgcactgag cacctggatt tataccatta ttcgtaacct gtgctttgac
     1081 ctgctgcgta aacagaaagg taaagaactg catattcaca gcgacgatat ttggccgagc
     1141 gaatattatc cgcctgatat ggttgatcat tatagtccgg aacaggatat gctgaaagaa
     1201 caggtggtta aatttctgga tatcctgccg aaaaatcagc gtgatgttct gcaagcagtt
     1261 tatctggaag aactgccgca tcagcaggtt gcagaactgt ttgatattcc gctgggcacc
     1321 gttaaaagcc gtctgcgtct ggcagttgaa aaactgcgtc atagcatgca taccgaacag
     1381 ctgtaataat actagagcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     1441 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     1501 ttcgggtggg cctttctgcg tttatatact agaggcctcc acaccgctcg tcacatcctg
     1561 tgatccactc ttcatcccgc tacgtaacac ctcttactag agattaaaga ggagaaatac
     1621 tagatgcgta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
     1681 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
     1741 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca
     1801 cttgtcacta ctttcggtta tggtgttcaa tgctttgcga gatacccaga tcatatgaaa
     1861 cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
     1921 ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt
     1981 gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac
     2041 aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat
     2101 ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca
     2161 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
     2221 tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc
     2281 cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaataa
     2341 taatactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2401 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     2461 tgggcctttc tgcgtttata tactagtagc ggccgctgca ggcatgcaag cttgcggccg
     2521 cgtcgtgact gggaaaaccc tggcgactag tcttggactc ctgttgatag atccagtaat
     2581 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     2641 tggtgagaat ccaggggtcc ccaataatta cgatttaaat tagtagcccg cctaatgagc
     2701 gggctttttt ttaattcccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2761 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagc
     2821 attcagcatt ttcgtgtggc gctgattccg ttttttgcgg cgttttgcct gccggtgttt
     2881 gcgcatccgg aaaccctggt gaaagtgaaa gatgcggaag atcaactggg tgcgcgcgtg
     2941 ggctatattg aactggatct gaacagcggc aaaattctgg aatcttttcg tccggaagaa
     3001 cgttttccga tgatgagcac ctttaaagtg ctgctgtgcg gtgcggttct gagccgtgtg
     3061 gatgcgggcc aggaacaact gggccgtcgt attcattata gccagaacga tctggtggaa
     3121 tatagcccgg tgaccgaaaa acatctgacc gatggcatga ccgtgcgtga actgtgcagc
     3181 gcggcgatta ccatgagcga taacaccgcg gcgaacctgc tgctgacgac cattggcggt
     3241 ccgaaagaac tgaccgcgtt tctgcataac atgggcgatc atgtgacccg tctggatcgt
     3301 tgggaaccgg aactgaacga agcgattccg aacgatgaac gtgataccac catgccggca
     3361 gcaatggcga ccaccctgcg taaactgctg acgggtgagc tgctgaccct ggcaagccgc
     3421 cagcaactga ttgattggat ggaagcggat aaagtggcgg gtccgctgct gcgtagcgcg
     3481 ctgccggctg gctggtttat tgcggataaa agcggtgcgg gcgaacgtgg cagccgtggc
     3541 attattgcgg cgctgggccc ggatggtaaa ccgagccgta ttgtggtgat ttataccacc
     3601 ggcagccagg cgacgatgga tgaacgtaac cgtcagattg cggaaattgg cgcgagcctg
     3661 attaaacatt ggtaaaccga tacaattaaa ggctcctttt ggagcctttt tttttggacg
     3721 acccttgtcc ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
     3781 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
     3841 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
     3901 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
     3961 gcgaggctgg ccgtaggccg gccgcgatgc aggtggctgc tgaaccccca gccggaactg
     4021 accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca
     4081 ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca
     4141 cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct
     4201 cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact
     4261 tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga
     4321 tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca
     4381 gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta
     4441 ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt
     4501 cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt
     4561 actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt
     4621 aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg
     4681 ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc
     4741 gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct
     4801 gcttcgtgtg tttcagcaac gcggcctgct tggcttcgct gacctgtttt gccaggtcct
     4861 cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact
     4921 tcgccaaacc tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg
     4981 cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct
     5041 ggactatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg
     5101 cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct
     5161 tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccggaa ttaattcccc
     5221 ggatcgatcc gtcgatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
     5281 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
     5341 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc
     5401 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     5461 catccggggt cagcaccgtt tctgcggact ggctttctac gtggctgcca tttttggggt
     5521 gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg cgttagcggg
     5581 ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg
     5641 cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt taaaagacag
     5701 gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg
     5761 acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg
     5821 tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg
     5881 cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat caggcgtttt
     5941 cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc tgcccctcat
     6001 ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt
     6061 cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccctac atggctctgc
     6121 tgtagtgagt gggttgcgct ccggcagcgg tcctgatccc ccgcagaaaa aaaggatctc
     6181 aagaagatcc tttgatcttt tctacggcgc gcccagctgt ctagggcggc ggatttgtcc
     6241 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     6301 gagcctttcg ttttatttga tgcct
//
