LOCUS       sequence-113925-         903 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PITCh donor vector for EGFP-2A-PuroR knock-in into human FBL locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 903)
  AUTHORS   Sakuma T, Nakade S, Sakane Y, Suzuki KT, Yamamoto T
  TITLE     MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the 
            PITCh systems.
  JOURNAL   Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub
            2015 Dec 17.
  PUBMED    26678082
REFERENCE   2  (bases 1 to 903)
  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/nprot.2015.140"; journalName: "Nat Protoc"; date: 
            "2016-01"; volume: "11"; issue: "1"; pages: "118-33"
COMMENT     Sequence Label: pCRIS-PITChv2-FBL
FEATURES             Location/Qualifiers
     source          1..903
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(33..743,744..746,747..749))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(33..743,744..746,747..749))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(75..96)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     424..443
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     683..704
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 gttagcagac ttcctctgcc ctctccgctt cccttgtaca gctcgtccat gccgagagtg
       61 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
      121 ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
      181 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
      241 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
      301 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
      361 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
      421 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
      481 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtaacg gctgaagcac
      541 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
      601 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
      661 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
      721 gtgaacagct cctcgccctt gctcaccatg gatccggggg gtggcctgtg agaggaacca
      781 aacacgtacg cgtacgatgc tatgtaacgc ggaactccat atatgggcta tgaactaatg
      841 accccgtaat tgattactat taataactag tcaataatca atgtcaacgc cactgtgcat
      901 gaa
//
