LOCUS       sequence-121735-        7297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALE for mouse major satellite with FLAG tag under CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7297)
  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 7297)
  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-MCS
FEATURES             Location/Qualifiers
     source          1..7297
                     /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             4297..4326
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4342..4359
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4382..4399)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4388..4612
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4622..4639)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4622..4638)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4631..4653)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4780..5235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4867..4886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5077..5098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5261..5365
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5366..5434,5435..6226)
                     /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(5584..5603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6397..6985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6886..6905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7139..7156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7273..7294
                     /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 gaattcgata
     4081 tcggcggccg cgtcgactga taattctgta catatgggca ccaaagaacc tgtaaacgtt
     4141 atctttttaa attgaatgtg cacaaataaa agtttggaaa agaaaaaaaa aaaaaaaaaa
     4201 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa atacgcatca
     4261 tacgctgtgg cgtacctcga gtctagaggg cccttcgaac aaaaactcat ctcagaagag
     4321 gatctgaata tgcataccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     4381 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4441 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4501 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4561 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat gggcggccgc
     4621 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4681 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4741 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     4801 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4861 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4921 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4981 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5041 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5101 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5161 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5221 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5281 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5341 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5401 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5461 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5521 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5581 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5641 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5701 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5761 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5821 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5881 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5941 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6001 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6061 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6121 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6181 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6241 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6301 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6541 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7021 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7081 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7141 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7201 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     7261 gcgcaacgca attaatgtga gttagctcac tcattag
//
