LOCUS       sequence-121734-        7447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALE for mouse major satellite with VP64 and FLAG under 
            CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7447)
  AUTHORS   Ishiuchi T, Enriquez-Gasca R, Mizutani E, Boskovic A, 
            Ziegler-Birling C, Rodriguez-Terrones D, Wakayama T, Vaquerizas JM, 
            Torres-Padilla ME
  TITLE     Early embryonic-like cells are induced by downregulating 
            replication-dependent chromatin assembly.
  JOURNAL   Nat Struct Mol Biol. 2015 Aug 3. doi: 10.1038/nsmb.3066.
  PUBMED    26237512
REFERENCE   2  (bases 1 to 7447)
  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 Struct 
            Mol Biol. 2015 Aug 3. doi: 10.1038/nsmb.3066."
COMMENT     Sequence Label: pTAL-MajSat-FLAG-NLS-VP64
FEATURES             Location/Qualifiers
     source          1..7447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(12..17,18..35,36..42)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    50..66
                     /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     55..77
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     74..90
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     74..90
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        110..489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        491..768
                     /label=chicken beta-actin promoter
     intron          769..1777
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1701..1723
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1790..1809
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1790..1808
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           1833..1875
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1896..1916
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4014..4037
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4041..4061
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4086..4235
                     /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             4447..4476
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4492..4509
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4532..4549)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4538..4762
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4772..4789)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4772..4788)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4781..4803)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4930..5385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5017..5036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5227..5248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5411..5515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5516..5584,5585..6376)
                     /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"
     primer_bind     complement(5734..5753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6547..7135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7036..7055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7289..7306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7423..7444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
       61 taacaatttc acacaggaaa cagctatgac catgaggcgc gccggattcg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      541 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      601 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      661 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      721 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      781 ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg
      841 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      901 cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct taaagggctc
      961 cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
     1021 ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1081 gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     1141 gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     1201 agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct ccccgagttg
     1261 ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg
     1321 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     1381 gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc
     1441 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     1501 ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     1561 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     1621 gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg gctgccttcg
     1681 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1741 tgctaaccat gttcatgcct tcttcttttt cctacagatc cttaattaat aatacgactc
     1801 actataggga acaaaagctg gagctaagct tgcttgttct ttttgcagaa gctcagaata
     1861 aacgctcaac tttggcagcc gccaccatga gatctcctaa gaaaaagagg aaggtgcagg
     1921 tggatctacg cacgctcggc tacagtcagc agcagcaaga gaagatcaaa ccgaaggtgc
     1981 gttcgacagt ggcgcagcac cacgaggcac tggtgggcca tgggtttaca cacgcgcaca
     2041 tcgttgcgct cagccaacac ccggcagcgt tagggaccgt cgctgtcacg tatcagcaca
     2101 taatcacggc gttgccagag gcgacacacg aagacatcgt tggcgtcggc aaacagtggt
     2161 ccggcgcacg cgccctggag gccttgctca cggatgcggg ggagttgaga ggtccgccgt
     2221 tacagttgga cacaggccaa cttgtgaaga ttgcaaaacg tggcggcgtg accgcaatgg
     2281 aggcagtgca tgcatcgcgc aatgcactga cgggtgcccc cctgaacctg accccggacc
     2341 aagtggtggc tatcgccagc aacggtggcg gcaagcaagc gctcgaaacg gtgcagcggc
     2401 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
     2461 gcaacaaggg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     2521 aggaccatgg cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc
     2581 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc
     2641 cggaccaagt ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc
     2701 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
     2761 tcgccagcaa cattggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     2821 tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc aacggtggcg
     2881 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     2941 tgaccccgga ccaagtggtg gctatcgcca gcaacattgg cggcaagcaa gcgctcgaaa
     3001 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
     3061 tggctatcgc cagcaacggt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
     3121 cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg
     3181 gtggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
     3241 atggcctgac tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc
     3301 tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc
     3361 aagtggtggc tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc
     3421 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
     3481 gcaacattgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     3541 aggaccatgg cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc
     3601 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
     3661 cggaccaagt ggtggctatc gccagcaaca agggcggcaa gcaagcgctc gaaacggtgc
     3721 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
     3781 tcgccagcaa cggtggcggc aagcaagcgc tcgaaagcat tgtggcccag ctgagccggc
     3841 ctgatccggc gttggccgcg ttgaccaacg accacctcgt cgccttggcc tgcctcggcg
     3901 gacgtcctgc catggatgca gtgaaaaagg gattgccgca cgcgccggaa ttgatcagaa
     3961 gagtcaatcg ccgtattggc gaacgcacgt cccatcgcgt tgcgggatcc atggactaca
     4021 aggacgacga tgacaagagc cccaagaaga agagaaaggt ggaggccagc ggttccggac
     4081 gggctgacgc attggacgat tttgatctgg atatgctggg aagtgacgcc ctcgatgatt
     4141 ttgaccttga catgcttggt tcggatgccc ttgatgactt tgacctcgac atgctcggca
     4201 gtgacgccct tgatgatttc gacctggaca tgctgattaa ctctagttga gaattctgta
     4261 catatgggca ccaaagaacc tgtaaacgtt atctttttaa attgaatgtg cacaaataaa
     4321 agtttggaaa agaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4381 aaaaaaaaaa aaaaaaaaaa atacgcatca tacgctgtgg cgtacctcga gtctagaggg
     4441 cccttcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg tcatcatcac
     4501 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4561 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4621 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4681 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4741 ggggatgcgg tgggctctat gggcggccgc cactggccgt cgttttacaa cgtcgtgact
     4801 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4861 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4921 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4981 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5041 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5101 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5161 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5221 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5281 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5341 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     5401 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5461 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5521 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5581 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5641 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5701 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5761 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5821 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5881 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5941 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6001 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6061 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6661 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7201 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7261 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7321 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7381 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7441 tcattag
//
