LOCUS       sequence-317150-        3405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5 repeats of Ptet2 sponge.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3405)
  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 3405)
  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: pXWS5Ptet2
FEATURES             Location/Qualifiers
     source          1..3405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(141..956)
                     /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(141..956)
                     /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     864..883
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     948..967
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(1551..2095)
                     /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(1630..1649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2563..2581
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2585..2628)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2631..2652
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    2693..2711
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2718..2736
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2813..2831
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2838..2856
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2933..2951
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2958..2976
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3053..3071
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3078..3096
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3173..3191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3198..3216
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      3265..3336
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3352..3379
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    join(3386..3405,1)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
       61 tcactcaaag gcggtaatct cgagtcccgt caagtcagcg taatgctctg ccagtgttac
      121 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
      181 ttcatatcag gattatcgat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
      241 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
      301 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
      361 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
      421 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
      481 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
      541 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
      601 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
      661 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
      721 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
      781 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
      841 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
      901 atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca
      961 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     1021 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     1081 acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg
     1141 gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt
     1201 ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca
     1261 ccttcttcac gaggcagacc tcagcgctag cggagtgtat actggcttac tatgttggca
     1321 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     1381 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     1441 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     1501 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     1561 ctccgccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     1621 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     1681 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     1741 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     1801 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     1861 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     1921 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     1981 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcaaaaaac cgccctgcaa
     2041 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     2101 tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattccat ggtgccacct
     2521 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2581 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg
     2641 ccgcttctag agatcgattt tagagctgcc ttttcagcag gacgcactga cctccctatc
     2701 agtgatagag attgacatcc ctatcagtga tagagatact gagcacatat cttaggtgtc
     2761 cctagaatga gtatcgattt tagagctgcc ttttcagcag gacgcactga cctccctatc
     2821 agtgatagag attgacatcc ctatcagtga tagagatact gagcacatat cttaggtgtc
     2881 cctagaattt ccatcgattt tagagctgcc ttttcagcag gacgcactga cctccctatc
     2941 agtgatagag attgacatcc ctatcagtga tagagatact gagcacatat cttaggtgtc
     3001 cctagaatga ctatcgattt tagagctgcc ttttcagcag gacgcactga cctccctatc
     3061 agtgatagag attgacatcc ctatcagtga tagagatact gagcacatat cttaggtgtc
     3121 cctagaattg atatcgattt tagagctgcc ttttcagcag gacgcactga cctccctatc
     3181 agtgatagag attgacatcc ctatcagtga tagagatact gagcacatat cttaggtgtc
     3241 cctagaatta ctagagccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     3301 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
     3361 tcgggtgggc ctttctgcgt ttatatacta gtagcggccg ctgca
//
