LOCUS       sequence-99974-e         958 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Part of Multibow set of constructs--each FP gene is initially not 
            expressed and then adopts a permanent ON or OFF status upon 
            Cre-mediated recombination. Encodes membrane bound EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 958)
  AUTHORS   Xiong F, Obholzer ND, Noche RR, Megason SG
  TITLE     Multibow: digital spectral barcodes for cell tracing.
  JOURNAL   PLoS One. 2015 May 26;10(5):e0127822. doi: 
            10.1371/journal.pone.0127822. eCollection 2015.
  PUBMED    26010570
REFERENCE   2  (bases 1 to 958)
  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: "PLoS One.";
            date: "2015-05-26"; volume: "10(5):e0127822. doi"; pages: " 
            10.1371/journal.pone.0127822. eCollection 2015"
COMMENT     Sequence Label: pMTB-Multibow-mG
FEATURES             Location/Qualifiers
     source          1..958
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(10..43)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      162..171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(203..205,206..916)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(203..205,206..916)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(245..266)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(506..525)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ggatctagga taacttcgta taggatactt tatacgaagt tattgatcaa ttcacagcca
       61 ccatgctgtg ctgcatcaga agaactaaac cggttgagaa gaatgaagag gccgatcagg
      121 agctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg ggagcgccag
      181 ctggaggtgc aggtgcagca aggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      241 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      301 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      361 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      421 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      481 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      541 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      601 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
      661 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
      721 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
      781 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
      841 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
      901 ggacgagctg tacaaggcag cagcatgata aatggtcctg ctggagttcg tgaccgcc
//
