LOCUS       sequence-318580-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AHL-responsive single-layered circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  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 5853)
  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: pXW101Plux2-gfp
FEATURES             Location/Qualifiers
     source          1..5853
                     /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_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             124..135
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      939..1010
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1026..1053
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1134..1145
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1152..1865
                     /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(1182..1210)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1805..1825
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1888..1959
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1975..2002
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2009..2029
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2084..2178
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        2251..2342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2343..2411,2412..3203)
                     /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      3207..3246
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            3393..3501
                     /note="incP origin of transfer"
     CDS             complement(3562..4710)
                     /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      5103..5634
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5761..5847)
                     /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 tattatgcta gctactagag
      121 attaaagagg agaaatacta gatgaaaaac ataaatgccg acgacacata cagaataatt
      181 aataaaatta aagcttgtag aagcaataat gatattaatc aatgcttatc tgatatgact
      241 aaaatggtac attgtgaata ttatttactc gcgatcattt atcctcattc tatggttaaa
      301 tctgatattt caatcctaga taattaccct aaaaaatgga ggcaatatta tgatgacgct
      361 aatttaataa aatatgatcc tatagtagat tattctaact ccaatcattc accaattaat
      421 tggaatatat ttgaaaacaa tgctgtaaat aaaaaatctc caaatgtaat taaagaagcg
      481 aaaacatcag gtcttatcac tgggtttagt ttccctattc atacggctaa caatggcttc
      541 ggaatgctta gttttgcaca ttcagaaaaa gacaactata tagatagttt atttttacat
      601 gcgtgtatga acataccatt aattgttcct tctctagttg ataattatcg aaaaataaat
      661 atagcaaata ataaatcaaa caacgattta accaaaagag aaaaagaatg tttagcgtgg
      721 gcatgcgaag gaaaaagctc ttgggatatt tcaaaaatat taggttgcag tgagcgtact
      781 gtcactttcc atttaaccaa tgcgcaaatg aaactcaata caacaaaccg ctgccaaagt
      841 atttctaaag caattttaac aggagcaatt gattgcccat actttaaaaa ttaataacac
      901 tgatagtgct agtgtagatc actactagag ccaggcatca aataaaacga aaggctcagt
      961 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     1021 acactggctc accttcgggt gggcctttct gcgtttatat actagagacc tgtaggatcg
     1081 tacaggttta cgcaagaaaa tggtttgtta ctttcgaata aatactagag attaaagagg
     1141 agaaatacta gatgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
     1201 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
     1261 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
     1321 ggccaacact tgtcactact ttcggttatg gtgttcaatg ctttgcgaga tacccagatc
     1381 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
     1441 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
     1501 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
     1561 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
     1621 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
     1681 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     1741 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc
     1801 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     1861 acaaataata atactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1921 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     1981 ccttcgggtg ggcctttctg cgtttatata ctagtagcgg ccgctgcagg catgcaagct
     2041 tgcggccgcg tcgtgactgg gaaaaccctg gcgactagtc ttggactcct gttgatagat
     2101 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     2161 ttttttattg gtgagaatcc aggggtcccc aataattacg atttaaatta gtagcccgcc
     2221 taatgagcgg gctttttttt aattccccta tttgtttatt tttctaaata cattcaaata
     2281 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2341 gtatgagcat tcagcatttt cgtgtggcgc tgattccgtt ttttgcggcg ttttgcctgc
     2401 cggtgtttgc gcatccggaa accctggtga aagtgaaaga tgcggaagat caactgggtg
     2461 cgcgcgtggg ctatattgaa ctggatctga acagcggcaa aattctggaa tcttttcgtc
     2521 cggaagaacg ttttccgatg atgagcacct ttaaagtgct gctgtgcggt gcggttctga
     2581 gccgtgtgga tgcgggccag gaacaactgg gccgtcgtat tcattatagc cagaacgatc
     2641 tggtggaata tagcccggtg accgaaaaac atctgaccga tggcatgacc gtgcgtgaac
     2701 tgtgcagcgc ggcgattacc atgagcgata acaccgcggc gaacctgctg ctgacgacca
     2761 ttggcggtcc gaaagaactg accgcgtttc tgcataacat gggcgatcat gtgacccgtc
     2821 tggatcgttg ggaaccggaa ctgaacgaag cgattccgaa cgatgaacgt gataccacca
     2881 tgccggcagc aatggcgacc accctgcgta aactgctgac gggtgagctg ctgaccctgg
     2941 caagccgcca gcaactgatt gattggatgg aagcggataa agtggcgggt ccgctgctgc
     3001 gtagcgcgct gccggctggc tggtttattg cggataaaag cggtgcgggc gaacgtggca
     3061 gccgtggcat tattgcggcg ctgggcccgg atggtaaacc gagccgtatt gtggtgattt
     3121 ataccaccgg cagccaggcg acgatggatg aacgtaaccg tcagattgcg gaaattggcg
     3181 cgagcctgat taaacattgg taaaccgata caattaaagg ctccttttgg agcctttttt
     3241 tttggacgac ccttgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     3301 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     3361 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     3421 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     3481 cctgctctgc gaggctggcc gtaggccggc cgcgatgcag gtggctgctg aacccccagc
     3541 cggaactgac cccacaaggc cctagcgttt gcaatgcacc aggtcatcat tgacccaggc
     3601 gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca
     3661 cttcttcacg cgggtggaat ccgatccgca catgaggcgg aaggtttcca gcttgagcgg
     3721 gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt
     3781 gcggtacttc tcccatatga atttcgtgta gtggtcgcca gcaaacagca cgacgatttc
     3841 ctcgtcgatc aggacctggc aacgggacgt tttcttgcca cggtccagga cgcggaagcg
     3901 gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt
     3961 cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa taccggccat tgatcgacca
     4021 gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg
     4081 cttcgcgtac tccaacacct gctgccacac cagttcgtca tcgtcggccc gcagctcgac
     4141 gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc
     4201 ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg
     4261 catcgctcgc atcgtgtccg gccacggcgc aatatcgaac aaggaaagct gcatttcctt
     4321 gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc
     4381 caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt
     4441 catcgacttc gccaaacctg ccgcctcctg ttcgagacga cgcgaacgct ccacggcggc
     4501 cgatggcgcg ggcagggcag ggggagccag ttgcacgctg tcgcgctcga tcttggccgt
     4561 agcttgctgg actatcgagc cgacggactg gaaggtttcg cggggcgcac gcatgacggt
     4621 gcggcttgcg atggtttcgg catcctcggc ggaaaacccc gcgtcgatca gttcttgcct
     4681 gtatgccttc cggtcaaacg tccgattcat tcaccctcct tgcgggattg ccccggaatt
     4741 aattccccgg atcgatccgt cgatcttgat cccctgcgcc atcagatcct tggcggcaag
     4801 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     4861 aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca
     4921 ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     4981 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt ggctgccatt
     5041 tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg
     5101 ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc
     5161 gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta
     5221 aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct
     5281 gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc
     5341 ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata
     5401 ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca
     5461 ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg
     5521 cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc
     5581 tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc ggccctacat
     5641 ggctctgctg tagtgagtgg gttgcgctcc ggcagcggtc ctgatccccc gcagaaaaaa
     5701 aggatctcaa gaagatcctt tgatcttttc tacggcgcgc ccagctgtct agggcggcgg
     5761 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     5821 tttcgactga gcctttcgtt ttatttgatg cct
//
