LOCUS       sequence-228223-        7834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor targeting vector for generating mouse Gatad2a conditional 
            knockout allele.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7834)
  AUTHORS   Mor N, Rais Y, Sheban D, Peles S, Aguilera-Castrejon A, Zviran A, 
            Elinger D, Viukov S, Geula S, Krupalnik V, Zerbib M, Chomsky E, 
            Lasman L, Shani T, Bayerl J, Gafni O, Hanna S, Buenrostro JD, Hagai 
            T, Masika H, Vainorius G, Bergman Y, Greenleaf WJ, Esteban MA, 
            Elling U, Levin Y, Massarwa R, Merbl Y, Novershtern N, Hanna JH
  TITLE     Neutralizing Gatad2a-Chd4-Mbd3/NuRD Complex Facilitates 
            Deterministic Induction of Naive Pluripotency.
  JOURNAL   Cell Stem Cell. 2018 Sep 6;23(3):412-425.e10. doi: 
            10.1016/j.stem.2018.07.004. Epub 2018 Aug 16.
  PUBMED    30122475
REFERENCE   2  (bases 1 to 7834)
  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: 
            "10.1016/j.stem.2018.07.004"; journalName: "Cell Stem Cell"; date: 
            "2018-09-6- 6"; volume: "23"; issue: "3"; pages: "412-425.e10"
COMMENT     Sequence Label: pNTK Gatad2a flox
FEATURES             Location/Qualifiers
     source          1..7834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(122..141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(801..1592,1593..1661))
                     /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     1424..1443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1662..1766)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      1810..1819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2908..2941
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2958..2991)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        3012..3511
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3047..3070)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3444..3463
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3525..4328
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3588..3607)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4198..4217
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4360..4377)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4366..4590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(4633..4666)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    4954..4987
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5790..5806)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6238..6286
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6311..6330
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6328..7458)
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMIQTHVTTPGSIPTICDLARTFAREMGEAN"
     promoter        complement(7537..7599)
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
ORIGIN
        1 gcaaaaggcc agaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1681 tattgaagca tttatcaggg ttattgtctc gtgagcggat acatatttga atgtatttag
     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcaaa
     1801 cagctatgac catggcggcc gcacctagtg actcacctca ccctgctcaa acccaaacca
     1861 cagagacaga tctgcactgt gacttccacc taactttatc ctgtgtcaca gtgccatcta
     1921 gtggacatat tctgagctgc tttccacaag caagttggta cttgcttggg tgctgggata
     1981 cacaccaagg ggcagtttag gttgtccact gtgagcagta ctttttctag gacttggtgc
     2041 tcctcccacc tccttcctgt gttacagcaa cagagcacac tggctttaag gatttagaag
     2101 caacagatgt gtatgccctc tgataccctt cacaaagagg gaggtctaat tttggggtgc
     2161 tcctgtgtgg gttaactgat gtaggctata ttatagccac tggtctttag ccagtgtggg
     2221 aagagtggtc agtgttgtgg acctgtgcat ctgacagtgt gtaccacccc tgctacaact
     2281 gagctcctag ggagactggc tgtttgctca cagggaaaag tgaacttcct acaccacagg
     2341 atgtgtgtta gaaggttggt cacagtggtg agtgttccat gatgtaactt cttctgagac
     2401 ttgagtgtgc agggttgagg gcagagccat cacactctgt agacctgagc atgaagtcag
     2461 tgagagtggt tcttttgagt ttgaaaactt gtagccactg cagccacttg ttatgtgtgt
     2521 cctaggtagg acaatgcagc agatggcccc aggtcccctt gtgaggtaga gagcaaaagc
     2581 taacagtaca tcattgtgct ttttggtctt ctggggggct gtatgggact gggttaagag
     2641 aggggtgggc ttcccattgg gtagttccca atgtgagacc ttggctttgg agtgctcacg
     2701 tccatcctac ccagcactca ggagctgaga tagcagacca agatctgaag ctctgtttac
     2761 ttgtgacaga gctctggggt cgagtagcac tttctgaagc atccatgcta cacctgagta
     2821 ctcagagatg gagcacacca tctggtgtgt atggccctaa tgaccagtga tgatacccat
     2881 ggtgcccacg cccgggtcta gccgcggata acttcgtata atgtatgcta tacgaagtta
     2941 tgaattctag gatcgatgaa gttcctatac tttctagaga ataggaactt cggatcccgg
     3001 cgcgccctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc
     3061 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc
     3121 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     3181 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     3241 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     3301 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag
     3361 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc
     3421 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct
     3481 gttctcctct tcctcatctc cgggcctttc gacctgcagc caatatggga tcggccattg
     3541 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3601 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3661 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3721 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3781 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3841 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3901 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3961 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4021 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4081 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4141 tttctggatt catcgactgt ggccggctgg gtgtggcgga tcgctatcag gacatagcgt
     4201 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4261 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4321 tcttctgagg ggatcaattc tctagagctc gctgatcagc ctcgactgtg ccttctagtt
     4381 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4441 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4501 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4561 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctgggggc
     4621 gcgcccctcg aggaagttcc tatactttct agagaatagg aacttcggta ccgtggcaag
     4681 aggaggcacc taggggtgat tgggcatctg gggtagtctg gcctgcctgg tactctgccc
     4741 tgctaagcac ctgcttatct ccatttgaca gtgacatgaa gtctgagaag agacctccct
     4801 cgcctgatgt gattgtgctc tctgacagtg agcagccatc gagcccaagg gtgaatggcc
     4861 tgaccacagt agccttgaaa gacaccagca ctgaggcgct cttggtgagc cttggcagag
     4921 cctggaagcc ccaccacccc accttgcaag cttataactt cgtataatgt atgctatacg
     4981 aagttataga tctagagctt ctgctgaggc tttccccaat tttgtgcttt tgctctccta
     5041 gaaaagcagc ccagaagaac gagagcggat gatcaagcag ctgaaggagg agctgaggct
     5101 agaggaagca aaactggtct tgttgaagaa gctgcggcag agtcagatac agaaggaagc
     5161 caccgcacag aaggtgcctg tgctctactg ccccagaccc cagagaggct ggcccagggc
     5221 agaggctcga cccacgggcg ggatggcagg cagtgtatgg cgttagggag caagcagaaa
     5281 gtaattcggg gtgggggggt gcgtgcgtgc gtgcgtgtaa ctttacacac caactccttt
     5341 tgatgtctct tgctatctta tagcctacag cttcctcagg gagcactgtg accacccctc
     5401 cccccctcgt gcggggtacc cagaacattc ctgctggcaa gacctcactt caggttcgtg
     5461 cgcccttctg cttgttgccc tgcaccacct ccgtttatag tcaggctcca aacactgcct
     5521 gctcctccca agtaaagaag catggggctc ccacagacta aaggaccaga gtgtcacatg
     5581 cgccccaaat tacaccacca aactaggcta ctatgaatgt cgcagactgg tcagtggtga
     5641 ttgagcagta agactgaaag attggctaga gtgggtcgct ccgctcaggc ctctgcccac
     5701 agactctggg gcctctgtca cactctcgta ttaccttcct ctgagttccc atggctcctg
     5761 ctagagtaat gggatctcct agagttaaca ctggccgtcg ttttaccggt ccgtagtcag
     5821 gtttagttgg tccggcggcg ccagaaatcc gcgcggtggt ttttgggggt cgggggtgtt
     5881 tggcagccac agacgcccgg tgttcgtgtc gcgccagtac atgcggtcca tgcccaggcc
     5941 atccaaaaac catgggtctg tctgctcagt ccagtcgtgg actgacccca cgcaacgccc
     6001 aaaataataa cccccacgaa ccataaacca ttccccatgg gggaccccgt ccctaaccca
     6061 cggggcccgt ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt
     6121 cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat
     6181 ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa
     6241 caaacgaccc aacacccgtg cgttttattc tgtcttttta ttgccgtcat agcgcaggtt
     6301 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc cccatctccc gtgcaaacgt
     6361 gcgcgccagg tcgcagatcg tcggtatgga gcctggggtg gtgacgtggg tctggatcat
     6421 cccggaggta agttgcagca gggcgtcccg gcagccggcg ggcgattggt cgtaatccag
     6481 gataaagacg tgcatgggac ggaggcgttt ggccaagacg tccaaggccc aggcaaacac
     6541 gttgtacagg tcgccgttgg gggccagcaa ctcgggggcc cgaaacaggg taaataacgt
     6601 gtccccgata tggggtcgtg ggcccgcgtt gctctggggc tcggcaccct ggggcggcac
     6661 ggccgtcccc gaaagctgtc cccaatcctc ccgccacgac ccgccgccct gcagataccg
     6721 caccgtattg gcaagcagcc cgtaaacgcg gcgaatcgcg gccagcatag ccaggtcaag
     6781 ccgctcgccg gggcgctggc gtttggccag gcggtcgatg tgtctgtcct ccggaagggc
     6841 ccccaacacg atgtttgtgc cgggcaaggt cggcgggatg agggccacga acgccagcac
     6901 ggcctggggg gtcatgctgc ccataaggta tcgcgcggcc gggtagcaca ggagggcggc
     6961 gatgggatgg cggtcgaaga tgagggtgag ggccgggggc ggggcatgtg agctcccagc
     7021 ctcccccccg atatgaggag ccagaacggc gtcggtcacg gcataaggca tgcccattgt
     7081 tatctgggcg cttgtcatta ccaccgccgc gtccccggcc gatatctcac cctggtcaag
     7141 gcggtgttgt gtggtgtaga tgttcgcgat tgtctcggaa gcccccagca cccgccagta
     7201 agtcatcggc tcgggtacgt agacgatatc gtcgcgcgaa cccagggcca ccagcagttg
     7261 cgtggtggtg gttttcccca tcccgtgggg accgtctata taaacccgca gtagcgtggg
     7321 cattttctgc tccgggcgga cttccgtggc ttcttgctgc cggcgagggc gcaacgccgt
     7381 acgtcggttg ctatggccgc gagaacgcgc agcctggtcg aacgcagacg cgtgctgatg
     7441 gccggggtac gaagccatac gcgcttctac aaggcgctgg ccgaagaggt gcgggagttt
     7501 cacgccacca agatgtgcgg cacgctgttg acgctgttaa gcgggtcgct gcagggtcgc
     7561 tcggtgttcg aggccacacg cgtcacctta atatgcgaag tggacctggg accgcgccgc
     7621 cccgactgca tctgcgtgtt ccaattcgcc aatgacaaga cgctgggcgg ggtttgctcg
     7681 acattgggtg gaaacattcc aggcctgggt ggagaggctt tttgcttcct cttgcaaaac
     7741 cacactgctc gacattgggt ggaaacattc caggcctggg tggagaggct ttttgcttcc
     7801 tcttgaaaac cacactgctc gactctacgg tccg
//
