LOCUS       sequence-442923-        1168 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  HITI insert construct with Ckm gRNA and GFP transgene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1168)
  AUTHORS   Padmaswari MH, Bulliard GN, Agrawal S, Jia MS, Nelson CE
  TITLE     Precision and efficacy of RNA-guided DNA integration in 
            high-expressing muscle loci.
  JOURNAL   bioRxiv [Preprint]. 2024 Mar 19:2024.03.18.582796. doi: 
            10.1101/2024.03.18.582796.
  PUBMED    38562818
REFERENCE   2  (bases 1 to 1168)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Mar 19:2024.03.18.582796. doi: 
            10.1101/2024.03.18.582796."
COMMENT     Sequence Label: p-HITI-GFP-Ckm
FEATURES             Location/Qualifiers
     source          1..1168
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      193..202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(199..201,202..204,205..918)
                     /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(199..201,202..204,205..915)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(244..265)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(505..524)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     852..873
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    945..1023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tgcgatncca ttaccctcac taaagggaac aaagctggga gctcccctat gtgatcctgc
       61 tggggactat gtccctaggt tcctggggag ggaggacgtc ggggccagcc aggcttaccc
      121 ccccctgccg ctgagacttc tgcgctgatg caccgcgcct cttcgcggtc tccctgtcct
      181 tgcagaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      241 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      301 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      361 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      421 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      481 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      541 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      601 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      661 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      721 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      781 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
      841 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      901 gacgagctgt acaagtaatc gtttagaccg ctgaaggatc cggcaacttg tttattgcag
      961 cttataatgg ttacaaataa agcaatagca tcacaatttc acaataaagc attttttcac
     1021 tgcattctag ttgtggnttg tccaactcat caatgtatct tagtcaggct gcccntccgc
     1081 aaaggnnccg gtcggggtcc cgcgtaacca cctggtgcac ccgaacgggc ttaacgggtg
     1141 gtgggtgtcc cgctctgcag gcgtctta
//
