LOCUS       sequence-59231-e        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Activation Domain: VP64; RVD of C-terminal 0.5 TALE repeat: NI 
            (binds A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Maeder ML, Linder SJ, Reyon D, Angstman JF, Fu Y, Sander JD, Joung 
            JK
  TITLE     Robust, synergistic regulation of human gene expression using TALE 
            activators.
  JOURNAL   Nat Methods. 2013 Feb 10. doi: 10.1038/nmeth.2366.
  PUBMED    23396285
REFERENCE   2  (bases 1 to 6122)
  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 
            Methods. 2013 Feb 10. doi: 10.1038/nmeth.2366."
COMMENT     Sequence Label: MLM3367
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     331..350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(569..673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(752..771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        927..2105
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1158..2096
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2053..2073
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2113..2122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2125..2145
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2971..3120
                     /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"
     primer_bind     complement(3162..3181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3216..3235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3272..3292)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3277..3606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3457..3592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3616..4215
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(3616..3635)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4112..4132
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4277..4318
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4328..4345
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4368..4385)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4374..4598
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4669..4685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4669..4685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4682..4704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4693..4709
                     /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)."
     promoter        complement(join(4717..4723,4724..4741,4742..4747))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4762..4783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4900..4917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5071..5659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5151..5170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5830..6122,1..499,500..568))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
       61 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
      241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
      301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
      361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
      421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
      481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
      541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
      601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
      721 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
      781 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      841 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      901 gttttgcgct gcttcgctag tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat
      961 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     1021 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     1081 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     1141 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     1201 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     1261 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     1321 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     1381 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     1441 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     1501 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     1561 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     1621 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     1681 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     1741 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     1801 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1861 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1921 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1981 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     2041 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     2101 cgtgaagcgg ccgccaccat ggcccccaag aagaagagga aggtgggcat tcaccgcggg
     2161 gtacctatgg tggacttgag gacactcggt tattcgcaac agcaacagga gaaaatcaag
     2221 cctaaggtca ggagcaccgt cgcgcaacac cacgaggcgc ttgtggggca tggcttcact
     2281 catgcgcata ttgtcgcgct ttcacagcac cctgcggcgc ttgggacggt ggctgtcaaa
     2341 taccaagata tgattgcggc cctgcccgaa gccacgcacg aggcaattgt aggggtcggt
     2401 aaacagtggt cgggagcgcg agcacttgag gcgctgctga ctgtggcggg tgagcttagg
     2461 gggcctccgc tccagctcga caccgggcag ctgctgaaga tcgcgaagag agggggagta
     2521 acagcggtag aggcagtgca cgcctggcgc aatgcgctca ccggggcccc cttgaacaga
     2581 gacgatctag agctagacgt ctcgctgaca cccgaacagg tggtcgccat tgcttctaac
     2641 atcggaggac ggccagcctt ggagagcatt gtggcccagc tgtccaggcc ggaccctgcc
     2701 ctggccgcgt taaccaacga ccacctcgtc gccttggcct gcctcggcgg acgtcctgcg
     2761 ctggatgcag tgaaaaaggg attgccgcac gcgccggcct tgatcaaaag aaccaatcgc
     2821 cgtattcccg aacgcacatc ccatcgcgtt gccgaccacg cgcaagtggt tcgcgtgctg
     2881 ggttttttcc agtgccactc ccacccagcg caagcatttg atgacgccat gacgcagttc
     2941 gggatgagcg gaggaggatc cgggcgcgcc gacgcgctgg acgatttcga tctcgacatg
     3001 ctgggttctg atgccctcga tgactttgac ctggatatgt tgggaagcga cgcattggat
     3061 gactttgatc tggacatgct cggctccgat gctctggacg atttcgatct cgatatgtta
     3121 taattaattg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3181 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3241 atcttatgaa ttccgattaa gtcgaccctg tggaatgtgt gtcagttagg gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3541 gcctcggctt ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3601 tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3661 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3721 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3781 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3841 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3901 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     3961 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtgtcgcc cgaccaccag
     4021 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     4081 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4141 ttcaccgtca ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc
     4201 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     4261 ccccatgcat ctcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     4321 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     4381 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4441 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4501 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4561 caatagcagg catgctgggg atgcggtggg ctccatggct tctgaggcgg aaagaaccag
     4621 ctggggctcg ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4681 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4741 gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4861 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5701 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5761 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5821 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5881 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5941 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6001 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6061 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6121 tt
//
