LOCUS       sequence-69391-e         785 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression plasmid for TALE proteins fused to VP64 activation domain
            compatible with GoldenGate cloning system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 785)
  AUTHORS   Perez-Pinera P, Ousterout DG, Brunger JM, Farin AM, Glass KA, Guilak
            F, Crawford GE, Hartemink AJ, Gersbach CA
  TITLE     Synergistic and tunable human gene activation by combinations of 
            synthetic transcription factors.
  JOURNAL   Nat Methods. 2013 Mar;10(3):239-42. doi: 10.1038/nmeth.2361. Epub 
            2013 Feb 3.
  PUBMED    23377379
REFERENCE   2  (bases 1 to 785)
  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: 
            "10.1038/nmeth.2361"; journalName: "Nat Methods"; date: "2013-03"; 
            volume: "10"; issue: "3"; pages: "239-42"
COMMENT     Sequence Label: pcDNA3.1-GoldenGate-VP64
FEATURES             Location/Qualifiers
     source          1..785
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(20..46)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(53..202)
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             complement(212..232)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    427..448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(463..468,469..486,487..493)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    501..517
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     506..528
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     525..541
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     525..541
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     560..580
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        562..580
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(627..646)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(628..646)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(656..673)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(656..672)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(665..687)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aattcctacc gattcaagaa gcgtagtccg gaacgtcgta cgggtagtta attaacatat
       61 cgagatcgaa atcgtccaga gcatcggagc cgagcatgtc cagatcaaag tcatccaatg
      121 cgtcgcttcc caacatatcc aggtcaaagt catcgagggc atcagaaccc agcatgtcga
      181 gatcgaaatc gtccagcgcg tcggcgcgcc caactttgcg tttctttttc gggctagcct
      241 tggatccggc aacgcgatgg gacgtgcgtt cgccaatacg gcgattgact cttctgatca
      301 attccggcgc gtgcggcaat ccctttttca ctgcatccat ggcaggacgt ccgccgaggc
      361 aggccaaggc gacgaggtgg tcgttggaga cgaccggtgg aaagcgggca gtgagcgcaa
      421 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      481 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      541 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggaggacct
      601 cgaggggggg cccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
      661 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      721 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      781 cccaa
//
