LOCUS       sequence-165859-        1059 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses optimized Sa sgCXCR4-3 gRNA with an EGFP fluorescent 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1059)
  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 1059)
  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: pSLQ2853-5 pHR: U6-Sasgv2CXCR4-3 CMV-EGFP
FEATURES             Location/Qualifiers
     source          1..1059
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    92..125
                     /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(145..858,859..861,862..864))
                     /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(190..211)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     539..558
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     798..819
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(898..997)
                     /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(927..947)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caatctttca caaattttgt aatccagagg ttgattatcg ataagcttga tatcgaattg
       61 tacctagtgg aaccggaacc cttaaacatg tataacttcg tataatgtat gctatacgaa
      121 gttattaggt ccctcgacga attcttactt gtacagctcg tccatgccga gagtgatccc
      181 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
      241 cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
      301 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
      361 gatcttgaag ttcgccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
      421 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
      481 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
      541 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
      601 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
      661 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
      721 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cagacacgct
      781 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
      841 cagctcctcg cccttgctca ccatggtggg ctagcggatc tgacggttca ctaaaccagc
      901 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
      961 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     1021 gtcaatgggg tggagacttg gaaatccccg tgagtcaaa
//
