LOCUS       sequence-422169-        4724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CombiGEM storage plasmid for ZF-2 CR fusions..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4724)
  AUTHORS   Alcantar MA, English MA, Valeri JA, Collins JJ
  TITLE     A high-throughput synthetic biology approach for studying 
            combinatorial chromatin-based transcriptional regulation.
  JOURNAL   Mol Cell. 2024 Jun 20;84(12):2382-2396.e9. doi: 
            10.1016/j.molcel.2024.05.025.
  PUBMED    38906116
REFERENCE   2  (bases 1 to 4724)
  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: 
            "10.1016/j.molcel.2024.05"; journalName: "Mol Cell"; date: 
            "2024-06-20- 20"; volume: "84"; issue: "12"; pages: "2382-2396.e9"
COMMENT     Sequence Label: pStorage_Ptet-2_ZF2
FEATURES             Location/Qualifiers
     source          1..4724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    131..149
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    170..188
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             212..277
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             284..304
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        1223..1410
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1411..2073
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     misc_feature    2288..2791
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(2808..3599)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4029..4617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4518..4537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaatta catctagact gaactggccg ataattgcag acgaacgctg aagatctagc
       61 tgccaaacca gatgtcaaca cagctacaac gctgctgttt ttttattttt tgttttttgc
      121 ggtataaaaa tccctatcag tgatagagat tttatttttt ttttcttttt ccctatcagt
      181 gatagagaat atcaagctac aaaaaggtat ggattataaa gaccatgacg gagattataa
      241 agatcatgac atagattaca aggatgacga tgacaagatg gcaccaaaga agaagaggaa
      301 ggtgggtatt cacggagtcc ccggaggtgg ctcagggtcc cgtcccggtg agaggccctt
      361 tcaatgcaga atttgcatgg caaacttttc cacggggcaa atcttggatc gtcatacaag
      421 aacgcatacg ggcgaaaaac cattccaatg ccgtatttgc atggctaact ttagcgttgc
      481 gcactccttg aagaggcacc tgagaacaca tactggcgag aagccctttc agtgcagaat
      541 ctgtatggcg aatttctccg atccctccaa tcttcgtaga catttgaaaa ctcatttaag
      601 aggatctggc ggtagtggcg gagggagtgg tggcgggggc tctggaggcg gatctgggtc
      661 ctgcaggggt tctgtcttct aataaataaa gcaatcttga tgaggataat gatttttttt
      721 tgaatataca taaatactac cgtttttctg ctagattttg tgatgacgta aataagtaca
      781 tattactttt taagccaaga caagattaag cattaacttt acccttttct ttctaagttt
      841 caatattagt tatcactgtt taaaagttat ggcgagaacg tcggcggtta aaatatatta
      901 ccctgaacgc ctcttccagc gctcaacggg tgtgcttccc gttctgatga gtccgtgagg
      961 acgaaagcgt cccgggaata tcgcgtagca ctttagactc ttccttacgt aatggatgac
     1021 tcggcgcgtg atcttacggc cgggcgacac ctctcatagg tgtgcgtgga tcctaacttt
     1081 tttggatccg caacggaatt cctgacgcac aattgttgtt gtttggctga gcagttacag
     1141 agatgttacg aaccactagt gcactgcagt acaaacacag tcctttcccg caattttctt
     1201 tttctattac tcttggcctc ctctagtaca ctctatattt ttttatgcct cggtaatgat
     1261 tttcattttt ttttttccac ctagcggatg actctttttt tttcttagcg attggcatta
     1321 tcacataatg aattatacat tatataaagt aatgtgattt cttcgaagaa tatactaaaa
     1381 aatgagcagg caagataaac gaaggcaaag atgacagagc agaaagccct agtaaagcgt
     1441 attacaaatg aaaccaagat tcagattgcg atctctttaa agggtggtcc cctagcgata
     1501 gagcactcga tcttcccaga aaaagaggca gaagcagtag cagaacaggc cacacaatcg
     1561 caagtgatta acgtccacac aggtataggg tttctggacc atatgataca tgctctggcc
     1621 aagcattccg gctggtcgct aatcgttgag tgcattggtg acttacacat agacgaccat
     1681 cacaccactg aggactgcgg gattgctctc ggtcaagctt ttaaagaggc cctaggggcc
     1741 gtgcgtggag taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga
     1801 gcggtggttg atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag
     1861 aaagtaggtg atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct
     1921 agcagaatta ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag
     1981 agtgcgttca aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac
     2041 gatgttccct ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgctgca
     2101 tacgatatat atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga
     2161 acagtatgat actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga
     2221 ggcgcgcttt ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc
     2281 atagagtatc acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa
     2341 attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat
     2401 gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct
     2461 caggtattaa tgccgaattg tttcatcttg tctgtgtaga aaaccacaca cgaaaatcct
     2521 gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg
     2581 tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt
     2641 tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac
     2701 aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata
     2761 cgaggcgcgt gtaagttaca ggcaagcgat ccgtcccgag cggccgctta tttgccgact
     2821 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
     2881 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
     2941 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
     3001 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
     3061 gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc
     3121 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
     3181 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
     3241 cccgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
     3301 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
     3361 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
     3421 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
     3481 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
     3541 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata
     3601 atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctccataac
     3661 atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt
     3721 accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg
     3781 cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagctta
     3841 cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccgtttcca cggtgtgcgt
     3901 cacccggcaa ccttgggtag cagcgaagtc gaggcatttc tgtcctggct ggtcatgacc
     3961 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4021 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4081 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4141 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4201 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4261 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4321 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4381 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4441 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4501 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4561 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4621 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4681 tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg ccgc
//
