LOCUS       sequence-138732-         876 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pFUGW-UBCp-RFP-CMVp-GFP-{BamHI+EcoRI}.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 876)
  AUTHORS   Wong AS, Choi GC, Cui CH, Pregernig G, Milani P, Adam M, Perli SD, 
            Kazer SW, Gaillard A, Hermann M, Shalek AK, Fraenkel E, Lu TK
  TITLE     Multiplexed barcoded CRISPR-Cas9 screening enabled by CombiGEM.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Feb 10. pii: 201517883.
  PUBMED    26864203
REFERENCE   2  (bases 1 to 876)
  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: "Proc Natl 
            Acad Sci U S A. 2016 Feb 10. pii: 201517883."
COMMENT     Sequence Label: pAWp9
FEATURES             Location/Qualifiers
     source          1..876
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(76..789,790..792,793..795))
                     /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             complement(join(79..789,790..792,793..795))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(121..142)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     470..489
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     729..750
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      792..801
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(814..865)
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(843..863)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 taatccagag gttgattatc gataagcttg atatcgaatt ccggactttt cgcggatcct
       61 agagtcgcgg ccgctttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
      121 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga
      181 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
      241 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
      301 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
      361 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
      421 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
      481 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
      541 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
      601 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
      661 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
      721 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
      781 gcccttgctc accatggtgg cgaccggtag gttaccgctg cttatataga cctcccaccg
      841 tacacgccta ccgcccattt gcgtcaatgg ggcgga
//
