LOCUS       sequence-211358-         717 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning backbone for sgRNA, co-expresses human optimized S. pyogenes
            Cas9 and ECFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 717)
  AUTHORS   Luo J, Lu L, Gu Y, Huang R, Gui L, Li S, Qi X, Zheng W, Chao T, 
            Zheng Q, Liang Y, Zhang L
  TITLE     Speed genome editing by transient CRISPR/Cas9 targeting and large 
            DNA fragment deletion.
  JOURNAL   J Biotechnol. 2018 Jun 7;281:11-20. doi: 
            10.1016/j.jbiotec.2018.06.308.
  PUBMED    29886029
REFERENCE   2  (bases 1 to 717)
  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: "J 
            Biotechnol."; date: "2018-06-7"; volume: "281:11-20. doi"; pages: " 
            10.1016/j.jbiotec.2018.06.308"
COMMENT     Sequence Label: pX458-ECFP
FEATURES             Location/Qualifiers
     source          1..717
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..3,4..717)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(43..64)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(304..323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     651..672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
       61 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      121 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      181 gtgaccaccc tgacctgggg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      241 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      301 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      361 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      421 ctggagtaca actacatcag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
      481 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
      541 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      601 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      661 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaa
//
