LOCUS       sequence-82029-e        8002 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  TALEN expressing vector used to allow Knock-in of OCT4-GFP-2A-PURO 
            knockin donor plasmid (generated by Jaenisch lab- Addgene Vector # 
            31939) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8002)
  AUTHORS   Gafni O, Weinberger L, Mansour AA, Manor YS, Chomsky E, Ben-Yosef D,
            Kalma Y, Viukov S, Maza I, Zviran A, Rais Y, Shipony Z, Mukamel Z, 
            Krupalnik V, Zerbib M, Geula S, Caspi I, Schneir D, Shwartz T, Gilad
            S, Amann-Zalcenstein D, Benjamin S, Amit I, Tanay A, Massarwa R, 
            Novershtern N, Hanna JH
  TITLE     Derivation of novel human ground state naive pluripotent stem cells.
  JOURNAL   Nature. 2013 Dec 12;504(7479):282-6. doi: 10.1038/nature12745. Epub 
            2013 Oct 30.
  PUBMED    24172903
REFERENCE   2  (bases 1 to 8002)
  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: 
            "10.1038/nature12745"; journalName: "Nature"; date: "2013-12-12- 
            12"; volume: "504"; issue: "7479"; pages: "282-6"
COMMENT     Sequence Label: REV human OCT4 TALEN
FEATURES             Location/Qualifiers
     source          1..8002
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        23..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..681
                     /label=chicken beta-actin promoter
     intron          683..1700
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1624..1646
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1708..1727
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1821..1841
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1842..1868
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             complement(2634..3041)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     2906..2922
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2932..2950
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2998..3018)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2998..3016)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3037..3053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3061..3077
                     /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(3085..3091,3092..3109,3110..3115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3130..3151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3847..4434
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     primer_bind     complement(4573..4592)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4638..4693
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4692..4711)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     regulatory      5059..5068
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5099..5117)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5124..5140)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(5281..5736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5418..5439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5630..5649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5815..5919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5920..5988,5989..6780)
                     /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(6138..6157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6951..7539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7440..7459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7693..7710
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7827..7848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7863..7868,7869..7886,7887..7893)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7901..7917
                     /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     7925..7941
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7959..7977
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7959..7976
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttgggagctc tcccatatgg tcgacattga ttattgacta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
      421 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      481 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg
      541 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
      601 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
      661 aagcgaagcg cgcggcgggc ggggagtcgc tgcgacgctg ccttcgcccc gtgccccgct
      721 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      781 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      841 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
      901 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
      961 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     1021 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     1081 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     1141 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     1201 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     1261 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
     1321 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     1381 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     1441 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     1501 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     1561 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     1621 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     1681 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     1741 ttggcaaaga attgatctaa ctcgagaaag atattgtata tatcgtaaca ataggaggtt
     1801 caacaatggc ttcctcccct ccaaagaaaa agagaaaggt tagttggaag gacgcaagtg
     1861 gttggtctag aatgcatgcg gatcccattc gtccgcgcag gccaagtcct gcccgcgagc
     1921 ttctgcccgg accccaaccg gatagggttc agccgactgc agatcgtggg gtgtctgcgc
     1981 ctgctggcag ccctctggat ggcttgcccg ctcggcggac ggtgtcccgg acccggctgc
     2041 catctccccc tgcgccctca cctgcgttct cggcgggcag cttcagcgat ctgctccgtc
     2101 cgttcgatcc gtcgcttctt gatacatcgc ttcttgattc gatgcctgcc gtcggcacgc
     2161 cgcatacagc ggctgcccca gcagagtggg atgaggcgca atcggctctg cgtgcagccg
     2221 atgacccgcc acccaccgtg cgtgtcgctg tcactgccgc gcggccgccg cgcgccaagc
     2281 cggccccgcg acggcgtgct gcgcaaccct ccgacgcttc gccggccgcg caggtggatc
     2341 tacgcacgct cggctacagt cagcagcagc aagagaagat caaaccgaag gtgcgttcga
     2401 cagtggcgca gcaccacgag gcactggtgg gccatgggtt tacacacgcg cacatcgttg
     2461 cgctcagcca acacccggca gcgttaggga ccgtcgctgt cacgtatcag cacataatca
     2521 cggcgttgcc agaggcgaca cacgaagaca tcgttggcgt cggcaaacag tggtccggcg
     2581 cacgcgccct ggaggccttg ctcacggatg cgggggagtt gagaggtccg ccgttacagt
     2641 tggacacagg ccaacttgtg aagattgcaa aacgtggcgg cgtgaccgca atggaggcag
     2701 tgcatgcatc gcgcaatgca ctgacgggtg cccccctgga gacgggcgcc gctacagggc
     2761 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     2821 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     2881 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
     2941 tcactatagg gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct
     3001 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3061 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3121 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3181 ccggtcgtct ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg tcctgccatg
     3241 gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt caatcgccgt
     3301 attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg cgtgctggag
     3361 tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac gcagttcggg
     3421 atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga actcgaagcc
     3481 cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca ggcatcaggg
     3541 atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc gtctttgcat
     3601 gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca cgagggagat
     3661 cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac cggcccctcc
     3721 gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca tttgcccctc
     3781 agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccgga tccgatatct
     3841 agatcccagc tagtgaaatc tgaattggaa gagaagaaat ctgaacttag acataaattg
     3901 aaatatgtgc cacatgaata tattgaattg attgaaatcg caagaaattc aactcaggat
     3961 agaatccttg aaatgaaggt gatggagttc tttatgaagg tttatggtta tcgtggtaaa
     4021 catttgggtg gatcaaggaa accagacgga gcaatttata ctgtcggatc tcctattgat
     4081 tacggtgtga tcgttgatac taaggcatat tcaggaggtt ataatcttcc aattggtcaa
     4141 gcagatgaaa tgcaaagata tgtcgaagag aatcaaacaa gaaacaagca tatcaaccct
     4201 aatgaatggt ggaaagtcta tccatcttca gtaacagaat ttaagttctt gtttgtgagt
     4261 ggtcatttca aaggaaacta caaagctcag cttacaagat tgaatcatat cactaattgt
     4321 aatggagctg ttcttagtgt agaagagctt ttgattggtg gagaaatgat taaagctggt
     4381 acattgacac ttgaggaagt gagaaggaaa tttaataacg gtgagataaa cttttaatag
     4441 gagctcgaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa aattcactcc
     4501 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4561 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4621 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4681 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4741 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     4801 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     4861 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     4921 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     4981 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc aggcggccgc
     5041 actagtgata tcccgcggcc atggcggccg ggagcatgcg acgtcgggcc caattcgccc
     5101 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5161 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5221 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5281 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     5341 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     5401 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5461 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5521 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5581 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5641 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5701 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcctg atgcggtatt ttctccttac
     5761 gcatctgtgc ggtatttcac accgcataca ggtggcactt ttcggggaaa tgtgcgcgga
     5821 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5881 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5941 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6001 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6061 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6121 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6181 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6241 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6301 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6361 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6421 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6481 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6541 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6601 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6661 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6721 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6781 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6841 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6901 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6961 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7021 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7081 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7141 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7201 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7261 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7321 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7381 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7441 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7501 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7561 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7621 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7681 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7741 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7801 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7861 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7921 cacacaggaa acagctatga ccatgattac gccaagctat ttaggtgaca ctatagaata
     7981 ctcaagctat gcatccaacg cg
//
