LOCUS       sequence-349752-        1405 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Targeting vector for knocking in constitutively expressed PE2-2A-GFP
            into the human AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1405)
  AUTHORS   Li H, Busquets O, Verma Y, Syed KM, Kutnowski N, Pangilinan GR, 
            Gilbert LA, Bateup HS, Rio DC, Hockemeyer D, Soldner F
  TITLE     Highly efficient generation of isogenic pluripotent stem cell models
            using prime editing.
  JOURNAL   Elife. 2022 Sep 7;11:e79208. doi: 10.7554/eLife.79208.
  PUBMED    36069759
REFERENCE   2  (bases 1 to 1405)
  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: "Elife."; 
            date: "2022-09-7"; volume: "11:e79208. doi"; pages: " 
            10.7554/eLife.79208"
COMMENT     Sequence Label: AAVS1-SA-neo-CAGGS-PE2-2A-GFP
FEATURES             Location/Qualifiers
     source          1..1405
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    13..238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     227..244
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(294..311)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(320..340)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(353..1063)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             complement(353..1063)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(395..416)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     744..763
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1003..1024
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     gap             1208..1217
                     /estimated_length=10
     gap             1233..1243
                     /estimated_length=11
     gap             1303..1335
                     /estimated_length=33
     gap             1345..1375
                     /estimated_length=31
     gap             1377..1404
                     /estimated_length=28
ORIGIN
        1 tccgctcaga agccatagna gcccaccgca tccccagcat gcctgctatt gtcttcccaa
       61 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
      121 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
      181 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc
      241 gaggctgatc agcgggttta aacgggccct ctagactcga ggaattaaac tcaatggtga
      301 tggtgatgat gaccggttag actttcctct tcttcttggg agaaccacca gacttgtaca
      361 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
      421 cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg
      481 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
      541 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
      601 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
      661 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
      721 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
      781 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
      841 cggggtagcg gctgaagcac tgcactccat aggtcagggt ggtcacgagg gtgggccagg
      901 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
      961 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     1021 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcccatag gtccagggtt
     1081 ctcctccncg tctccagcct gcttnagcag gctgaagttn nnagctccgc ttccgacttt
     1141 ccnnttctnn tgggctcnaa ttngctgccn nnggnggtnn ttttgagcng ccanaggntg
     1201 nnaatttnnn nnnnnnnggg aaanggnnnn ngnnnnnnnn nnngnnngcc tttcggnccc
     1261 tnggnnnnnn nccggnnncn nnnccccggg gnnnnnnnnn ttnnnnnnnn nnnnnnnnnn
     1321 nnnnnnnnnn nnnnnannnn nnntnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnntnnnn
     1381 nnnnnnnnnn nnnnnnnnnn nnnnt
//
