LOCUS       sequence-165853-        1026 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses optimized Sa sgCXCR4-2 gRNA with an EGFP fluorescent 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1026)
  AUTHORS   Gao Y, Xiong X, Wong S, Charles EJ, Lim WA, Qi LS
  TITLE     Complex transcriptional modulation with orthogonal and inducible 
            dCas9 regulators.
  JOURNAL   Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042.
  PUBMED    27776111
REFERENCE   2  (bases 1 to 1026)
  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. 2016 Oct 24. doi: 10.1038/nmeth.4042."
COMMENT     Sequence Label: pSLQ2853-4 pHR: U6-Sasgv2CXCR4-2 CMV-EGFP
FEATURES             Location/Qualifiers
     source          1..1026
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    93..126
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(146..859,860..862,863..865))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(191..212)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     540..559
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     799..820
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(899..998)
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(928..948)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tcaatctttc acaaattttg taatccagag gttgattatc gataagcttg atatcgaatt
       61 gtacctagtg gaaccggaac ccttaaacat gtataacttc gtataatgta tgctatacga
      121 agttattagg tccctcgacg aattcttact tgtacagctc gtccatgccg agagtgatcc
      181 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
      241 tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
      301 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
      361 ggatcttgaa gttcgccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
      421 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
      481 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
      541 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
      601 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
      661 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
      721 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccagacacgc
      781 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
      841 acagctcctc gcccttgctc accatggtgg gctagcggat ctgacggttc actaaaccag
      901 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
      961 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     1021 cgtcaa
//
