LOCUS       sequence-121267-        1091 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Integration-free (episomal) expression of EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1091)
  AUTHORS   Okita K, Matsumura Y, Sato Y, Okada A, Morizane A, Okamoto S, Hong 
            H, Nakagawa M, Tanabe K, Tezuka KI, Shibata T, Kunisada T, Takahashi
            M, Takahashi J, Saji H, Yamanaka S
  TITLE     A more efficient method to generate integration-free human iPS 
            cells.
  JOURNAL   Nat Methods. 2011 May;8(5):409-12
  PUBMED    21460823
REFERENCE   2  (bases 1 to 1091)
  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 
            Methods."; date: "2011-05"; volume: "8(5)"; pages: "409-1"
COMMENT     Sequence Label: pCXLE-EGFP
FEATURES             Location/Qualifiers
     source          1..1091
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     51..70
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      98..107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(104..106,107..109,110..823)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(104..106,107..109,110..820)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(149..170)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(410..429)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     757..778
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(908..928)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
       61 ggttattgtg ctgtctcatc attttggcaa agaattcgcc accatggtga gcaagggcga
      121 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      181 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      241 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
      301 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
      361 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      421 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      481 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      541 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
      601 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
      661 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
      721 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
      781 cgccgccggg atcactctcg gcatggacga gctgtacaag taagaattca agcttcgggg
      841 actagtcata tgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
      901 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
      961 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1021 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1081 acgcaacccc c
//
