LOCUS       sequence-165851-        1014 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses optimized Sa sgCXCR4-1 gRNA with an EGFP fluorescent 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1014)
  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 1014)
  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: pSLQ2853-3 pHR: U6-Sasgv2CXCR4-1 CMV-EGFP
FEATURES             Location/Qualifiers
     source          1..1014
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    94..127
                     /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(147..860,861..863,864..866))
                     /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(192..213)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     541..560
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     800..821
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(900..999)
                     /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(929..949)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtcaatcttt cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat
       61 tgtacctagt ggaaccggaa cccttaaaca tgtataactt cgtataatgt atgctatacg
      121 aagttattag gtccctcgac gaattcttac ttgtacagct cgtccatgcc gagagtgatc
      181 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
      241 ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
      301 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
      361 cggatcttga agttcgcctt gatgccgttc ttctgcttgt cggccatgat atagacgttg
      421 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
      481 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
      541 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
      601 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc
      661 acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
      721 cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccagacacg
      781 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
      841 aacagctcct cgcccttgct caccatggtg ggctagcgga tctgacggtt cactaaacca
      901 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
      961 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actc
//
