LOCUS       sequence-227769-        6966 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Non-replicating episomal expression of EBNA1, which enhances 
            transfection efficiency and expression from episomal plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6966)
  AUTHORS   Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A, 
            Goshima N, Yamanaka S
  TITLE     An Efficient Non-viral Method to Generate Integration-Free Human iPS
            Cells from Cord Blood and Peripheral Blood Cells.
  JOURNAL   Stem Cells. 2012 Nov 29. doi: 10.1002/stem.1293.
  PUBMED    23193063
REFERENCE   2  (bases 1 to 6966)
  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: "Stem Cells.
            2012 Nov 29. doi: 10.1002/stem.1293."
COMMENT     Sequence Label: pCXWB-EBNA1
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          103..1119
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1043..1065
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1127..1146
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1189..3114
                     /codon_start=1
                     /product="Epstein-Barr nuclear antigen 1, also known as 
                     EBNA-1"
                     /label=EBNA1
                     /note="crucial for latent viral infection, and for episomal
                     amplification of vectors containing the oriP origin (Okita 
                     et al., 2013)"
                     /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
                     GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
                     AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA
                     GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
                     GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG
                     GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     misc_feature    3158..3746
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3211..3231)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3629..3640)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3837..3856)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3902..3957
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3956..3975)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4318..4334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4318..4334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4331..4353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4342..4358
                     /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(4366..4372,4373..4390,4391..4396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4411..4432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4557..4574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4728..5316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4808..4827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5487..6278,6279..6347))
                     /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     6110..6129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6348..6452)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6483..6862
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
       61 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      121 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      181 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      241 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      301 gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
      361 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
      421 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
      481 ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
      541 gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct
      601 gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg
      661 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg
      721 gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg
      781 agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc
      841 caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
      901 gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg
      961 cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt
     1021 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     1081 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     1141 attgtgctgt ctcatcattt tggcaaagaa ttcgcccttg tcgacaccat gtctgacgag
     1201 gggccaggta caggacctgg aaatggccta ggagagaagg gagacacatc tggaccagaa
     1261 ggctccggcg gcagtggacc tcaaagaaga gggggtgata accatggacg aggacgggga
     1321 agaggacgag gacgaggagg cggaagacca ggagccccgg gcggctcagg atcagggcca
     1381 agacatagag atggtgtccg gagaccccaa aaacgtccaa gttgcattgg ctgcaaaggg
     1441 acccacggtg gaacaggagc aggagcagga gcgggagggg caggagcagg aggggcagga
     1501 gcaggaggag gggcaggagc aggaggaggg gcaggagggg caggaggggc aggaggggca
     1561 ggagcaggag gaggggcagg agcaggagga ggggcaggag gggcaggagg ggcaggagca
     1621 ggaggagggg caggagcagg aggaggggca ggaggggcag gagcaggagg aggggcagga
     1681 ggggcaggag gggcaggagc aggaggaggg gcaggagcag gaggaggggc aggaggggca
     1741 ggagcaggag gaggggcagg aggggcagga ggggcaggag caggaggagg ggcaggagca
     1801 ggaggggcag gaggggcagg aggggcagga gcaggagggg caggagcagg aggaggggca
     1861 ggaggggcag gaggggcagg agcaggaggg gcaggagcag gaggggcagg agcaggaggg
     1921 gcaggagcag gaggggcagg aggggcagga gcaggagggg caggaggggc aggagcagga
     1981 ggggcaggag gggcaggagc aggaggaggg gcaggagggg caggagcagg aggaggggca
     2041 ggaggggcag gagcaggagg ggcaggaggg gcaggagcag gaggggcagg aggggcagga
     2101 gcaggagggg caggaggggc aggagcagga ggaggggcag gagcaggagg ggcaggagca
     2161 ggaggtggag gccggggtcg aggaggcagt ggaggccggg gtcgaggagg tagtggaggc
     2221 cggggtcgag gaggtagtgg aggccgccgg ggtagaggac gtgaaagagc caggggggga
     2281 agtcgtgaaa gagccagggg gagaggtcgt ggacgtggag aaaagaggcc caggagtccc
     2341 agtagtcagt catcatcatc cgggtctcca ccgcgcaggc cccctccagg tagaaggcca
     2401 tttttccacc ctgtagggga agccgattat tttgaatacc accaagaagg tggcccagat
     2461 ggtgagcctg acgtgccccc gggagcgata gagcagggcc ccgcagatga cccaggagaa
     2521 ggcccaagca ctggaccccg gggtcagggt gatggaggca ggcgcaaaaa aggagggtgg
     2581 tttggaaagc atcgtggtca aggaggttcc aacccgaaat ttgagaacat tgcagaaggt
     2641 ttaagagctc tcctggctag gagtcacgta gaaaggacta ccgacgaagg aacttgggtc
     2701 gccggtgtgt tcgtatatgg aggtagtaag acctcccttt acaacctaag gcgaggaact
     2761 gcccttgcta ttccacaatg tcgtcttaca ccattgagtc gtctcccctt tggaatggcc
     2821 cctggacccg gcccacaacc tggcccgcta agggagtcca ttgtctgtta tttcatggtc
     2881 tttttacaaa ctcatatatt tgctgaggtt ttgaaggatg cgattaagga ccttgttatg
     2941 acaaagcccg ctcctacctg caatatcagg gtgactgtgt gcagctttga cgatggagta
     3001 gatttgcctc cctggtttcc acctatggtg gaaggggctg ccgcggaggg tgatgacgga
     3061 gatgacggag atgaaggagg tgatggagat gagggtgagg aagggcagga gtgaggatcc
     3121 aagggcgaat tcaagcttcg gggactagtc atatgataat caacctctgg attacaaaat
     3181 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3241 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3301 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3361 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3421 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3481 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3541 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3601 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3661 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3721 gatctccctt tgggccgcct ccccgcatcg gtacaattca ctcctcaggt gcaggctgcc
     3781 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     3841 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     3901 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3961 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     4021 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     4081 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     4141 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     4201 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     4261 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     4321 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4381 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gctgcattaa
     4501 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4561 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4621 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4681 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4741 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4801 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4861 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4921 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4981 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5041 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5101 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5161 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5221 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5281 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5341 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5401 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5461 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5521 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5581 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5641 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5701 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5761 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5821 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5881 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5941 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6001 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6061 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6121 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6181 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6241 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6301 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6361 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6421 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg
     6481 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     6541 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     6601 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     6661 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     6721 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     6781 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     6841 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
     6901 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
     6961 gatggg
//
