LOCUS       sequence-160939-        8295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FLAG-HA tagged human CBFA2T2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8295)
  AUTHORS   Tu S, Narendra V, Yamaji M, Vidal SE, Rojas LA, Wang X, Kim SY, 
            Garcia BA, Tuschl T, Stadtfeld M, Reinberg D
  TITLE     Co-repressor CBFA2T2 regulates pluripotency and germline 
            development.
  JOURNAL   Nature. 2016 Jun 8;534(7607):387-90. doi: 10.1038/nature18004.
  PUBMED    27281218
REFERENCE   2  (bases 1 to 8295)
  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"; journalName: 
            "Nature"; date: "2016-06-8- 8"; volume: "534"; issue: "7607"; pages:
            "387-90"
COMMENT     Sequence Label: CAGhCBFA2T2
FEATURES             Location/Qualifiers
     source          1..8295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    588..722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        736..1115
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1118..1393
                     /label=chicken beta-actin promoter
     intron          1394..2402
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2326..2348
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      2463..2472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2472..2495
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4308..4334
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(4371..4944,4945..4947,4948..4956)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4538..4555)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4765..4784
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4957..5556
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4957..4976)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5453..5473
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5637..5654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5643..5867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5914..5930)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5914..5930)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5927..5949)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5938..5954
                     /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(5962..5968,5969..5986,5987..5992))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6007..6028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6087..6282
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6133..6268
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6288..6422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6490..6507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6661..7249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6741..6760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7420..8211,8212..8280))
                     /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     8043..8062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atgaccgctt tggaaaaaca aagactgtat ttcctggaaa ttaatgttta ttcaataaac
       61 tgtgtattca gctatatcac atagtggtga ggctgaaatg aggcgggaag aggcggttgg
      121 ggcttaatta tatcaatttg ggtggcccca cagcgcctcc aaggcgccag tcctgttttg
      181 acaagttgcc tctggaagcc tctacaatgc ctctcttctt tttctccaga gtaagcggag
      241 gccaggggcc cccggcctct gcttaatact aaaaaaaaca gctgttgtca tagtaatgat
      301 tgggtggaaa cattccaggc ctgggtggag aggctttttg cttcctcttg caaaaccaca
      361 ctgacattcc aggcctgggt ggagaggctt tttgcttcct cttgcaaaac cacactgccc
      421 tctggagggc agttgcctag caactaacta aaagaggatg tcgcacggcc agctgcggtc
      481 agttagtcac ttcctgctta actgacttga cattttctat tttaagagtc gggaggaaaa
      541 ttactgtgtt ggaggccctc cgccatcttc tgaagctgaa tcgaattaac ttgtttattg
      601 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
      661 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
      721 tctgatatca tcgtcgacat tgattattga ctagttatta atagtaatca attacggggt
      781 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      841 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      901 taacgccaat agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc
      961 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1021 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1081 agtacatcta cgtattagtc atcgctatta ccatgggtcg aggtgagccc cacgttctgc
     1141 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     1201 ttattttgtg cagcgatggg ggcggggggg gggggggcgc gcgccaggcg gggcggggcg
     1261 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
     1321 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     1381 cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt gccccgctcc gcgccgcctc
     1441 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     1501 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
     1561 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
     1621 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggcccgcgc tgcccggcgg
     1681 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcgtgtgcg cgaggggagc
     1741 gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga ggggaacaaa ggctgcgtgc
     1801 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc ggcggtcggg ctgtaacccc
     1861 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1921 gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1981 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggccccgga
     2041 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     2101 agagggcgca gggacttcct ttgtcccaaa tctggcggag ccgaaatctg ggaggcgccg
     2161 ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     2221 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccatctcca gcctcggggc
     2281 tgccgcaggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     2341 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     2401 agctcctggg caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa gaattcctcg
     2461 aggccaccat ggactacaaa gacgatgacg acaagatggc taaagaatct ggaataagct
     2521 tgaaagaaat acaggtcctg gcaaggcaat ggaaagttgg tcctgagaaa agggtgccag
     2581 cgatgcctgg atcgcctgtg gaagtgaaga tacagtccag atcctcacct cccaccatgc
     2641 cacccctccc accaataaat cctggaggac cgaggccagt gtccttcact cctactgcat
     2701 taagcaatgg catcaaccat tctcctccta ccctgaatgg tgccccatca ccgccacaga
     2761 gattcagcaa tggtcctgcc tcctccacat catctgcact cacaaatcag caattgccag
     2821 ccacttgtgg tgctcgacaa ctcagcaagt tgaaacgctt tcttaccact ctgcaacagt
     2881 ttggcaatga catctcccct gagattgggg agaaggtgcg gactcttgtt cttgcactgg
     2941 tgaactcaac agtgacaatt gaggaattcc actgtaagct ccaagaagcc acaaactttc
     3001 cccttcgtcc ttttgtgatt ccatttctca aggccaacct gcccctgctg cagcgggaac
     3061 tgctgcactg cgctcgggcg gccaagcaga ccccatccca gtacctggct cagcacgaac
     3121 accttctgct caacacaagc attgcatcgc ctgctgactc gtcagagttg ctcatggagg
     3181 tgcacggaaa tgggaagagg cccagtccag agaggagaga agagaatagt tttgatagag
     3241 acacaattgc tcctgagcct cctgccaaga gagtatgtac catcagccct gctcctcggc
     3301 acagtcctgc tctcactgtg cccctcatga atcccggggg ccaattccat cctacccctc
     3361 cacctcttca gcattacacc ttagaggata ttgcaacttc tcacctgtat cgggaaccca
     3421 acaagatgct agagcatcga gaagttcgtg atagacacca cagtcttggt ctaaatggag
     3481 gctatcaaga tgagttggta gatcatcgtt tgacagaaag ggaatgggct gatgaatgga
     3541 aacatcttga ccatgcgctg aattgcatta tggaaatggt agagaaaaca aggcgctcta
     3601 tggcagttct gcggcgctgt caggaatcag atcgtgaaga actcaactac tggaaaagac
     3661 ggtacaatga aaacacagag ctgaggaaaa cggggaccga gttggtctcc aggcagcaca
     3721 gccctgggag tgcagattct ctcagcaatg attctcagag agagttcaac agcaggccag
     3781 gtacaggata cgtacctgtg gagttttgga aaaaaacaga agaagctgtg aataaggtga
     3841 aaattcaggc catgtcagaa gtacagaagg ccgtcgctga ggcagagcag aaagcctttg
     3901 aagtgattgc aacagagaga gcacgaatgg agcaaaccat agcggatgtc aagcggcagg
     3961 ccgcagagga tgctttcctc gtcatcaatg agcaagagga gtccacggag aactgctgga
     4021 actgtggccg caaagccagc gagacatgca gtggctgcaa tatcgcgcga tactgtggct
     4081 ctttctgcca gcacaaggac tgggagcggc accaccgcct ctgtggtcag aacctgcatg
     4141 gccagagccc ccacggccag ggccggccgc tgcttcctgt aggcaggggc tcctctgcca
     4201 ggtccgccga ctgcagcgtg cccagcccag ccctcgacaa gacctcggca accacatcgc
     4261 gttcctcaac acctgcttct gtgacagcta tcgacaccaa cggactctac ccatacgatg
     4321 ttccagatta cgcttaggcg gccgctcgat aagcttgata tcgaattccg cccctctccc
     4381 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     4441 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     4501 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     4561 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     4621 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     4681 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     4741 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     4801 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     4861 acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     4921 tcctttgaaa aacacgatga taatatggcc acaaccatga ccgagtacaa gcccacggtg
     4981 cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc
     5041 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     5101 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     5161 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     5221 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     5281 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     5341 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     5401 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     5461 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     5521 cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc
     5581 ccgaccgaaa ggagcgcacg accccatgca tcgatgatct agagctcgct gatcagcctc
     5641 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5701 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5761 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5821 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     5881 aagaacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5941 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6001 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6061 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     6121 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6181 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6241 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     6301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     6421 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6481 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6841 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7081 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8281 actcttcctt tttca
//
