LOCUS       sequence-311319-        1780 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  aTc-responsive single-layered circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1780)
  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 1780)
  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: pXW117Ptet2-gfp
FEATURES             Location/Qualifiers
     source          1..1780
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        1..35
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             47..58
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             65..688
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(138..157)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      708..779
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      795..822
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    837..855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    862..880
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             906..917
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             924..1637
                     /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(954..982)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1577..1597
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1660..1731
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1747..1774
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ttgacagcta gctcagtcct agggattgtg ctagctacta gagattaaag aggagaaata
       61 ccatatgtcc agattagata aaagtaaagt gattaacagc gcattagagc tgcttaatga
      121 ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc
      181 tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat
      241 gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt
      301 acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt
      361 acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt
      421 tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg ctgtggggca
      481 ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga
      541 aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca
      601 ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatttgcg gattagaaaa
      661 acaacttaaa tgtgaaagtg ggtcctaata atactagagc caggcatcaa ataaaacgaa
      721 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
      781 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagtccc
      841 tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac atattactag
      901 agattaaaga ggagaaatac tagatgcgta aaggagaaga acttttcact ggagttgtcc
      961 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
     1021 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
     1081 tacctgttcc atggccaaca cttgtcacta ctttcggtta tggtgttcaa tgctttgcga
     1141 gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg
     1201 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
     1261 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
     1321 atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca
     1381 tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag
     1441 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     1501 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
     1561 aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca
     1621 tggatgaact atacaaataa taatactaga gccaggcatc aaataaaacg aaaggctcag
     1681 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     1741 cacactggct caccttcggg tgggcctttc tgcgtttata
//
