LOCUS       sequence-199892-        3781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express two TALE-VP64 activator linked by in-frame T2A (encoding 
            Thosea asigna 'self-cleaving' 2A peptide) sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3781)
  AUTHORS   Lowder LG, Zhou J, Zhang Y, Malzahn A, Zhong Z, Hsieh TF, Voytas DF,
            Zhang Y, Qi Y
  TITLE     Robust transcriptional activation in plants using multiplexed 
            CRISPR-Act2.0 and mTALE-Act systems.
  JOURNAL   Mol Plant. 2017 Nov 29. pii: S1674-2052(17)30344-1. doi: 
            10.1016/j.molp.2017.11.010.
  PUBMED    29197638
REFERENCE   2  (bases 1 to 3781)
  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: "Mol Plant. 
            2017 Nov 29. pii: S1674-2052(17)30344-1. doi: 
            10.1016/j.molp.2017.11.010."
COMMENT     Sequence Label: pYPQ121
FEATURES             Location/Qualifiers
     source          1..3781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             27..50
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             54..74
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             182..202
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             335..388
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             395..421
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             422..442
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             483..503
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      649..901
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    946..1069
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1086..1105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1087..1105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1110..1126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1110..1126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1557..2348
                     /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      2441..3029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2930..2949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3183..3200
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3359..3386)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3478..3564)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3613..3635
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3627..3644
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3628..3644
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3660..3759
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gcgaagcttg gcttggtacc gatatggact acaaagacga tgacgataaa gctcctaaga
       61 aaaagcgcaa agtcccctct agataacgca ggatccgtaa gtttatcagt taaatataat
      121 aaataaagaa gaaaaccaaa aaaatggcta actaaaacga tggtcttatg attttatgca
      181 gccaaaaaag aaacgtaagg ttgatgctct tgacgatttt gacctcgata tgctcgacgc
      241 tcttgatgat tttgatctcg acatgctcga tgcacttgat gactttgacc ttgacatgct
      301 cgacgcactc gatgacttcg acctcgacat gcttgagggc agaggaagtc ttctaacatg
      361 cggtgacgtg gaggagaatc ccggccctat cgattaccct tatgatgtgc cagactacgc
      421 tcctaaaaag aagcgcaaag ttggcattca cggggtgccg gctagctaat aggcttagat
      481 ctccaaaaaa gaaacgtaag gttgatgctc ttgacgattt tgacctcgat atgctcgacg
      541 ctcttgatga ttttgatctc gacatgctcg atgcacttga tgactttgac cttgacatgc
      601 tcgacgcact cgatgacttc gacctcgaca tgctttaata ggacgtccga tcgttcaaac
      661 atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata
      721 taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt
      781 atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac
      841 aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat
      901 cgggaattga tcccccctcg acagcttccg gaaagggcga attcgcaact ttgtatacaa
      961 aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     1021 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgc catccagctg
     1081 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1141 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     1201 ggaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     1261 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     1321 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1381 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     1441 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     1501 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacatcatga
     1561 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     1621 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     1681 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     1741 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     1801 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     1861 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     1921 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     1981 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     2041 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     2101 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     2161 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     2221 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     2281 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     2341 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     2401 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2461 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2521 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2581 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2641 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2701 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2761 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2821 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2881 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2941 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3001 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3061 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3121 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3181 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3241 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3301 gtgagcgcaa cgcaattaat acgcgtaccg cgagccagga agagtttgta gaaacgcaaa
     3361 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     3421 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     3481 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     3541 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgcttg
     3601 catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag ctcgggcccc
     3661 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat
     3721 gcttttttat aatgccaact ttgtacaaaa aagcaggctc cgaattcgcc cttcaccatg
     3781 g
//
