LOCUS       sequence-267746-        4200 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses fluorescent ALIX S718/721A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4200)
  TITLE     Plasmids for creating inducible cell lines expressing ALIX-GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4200)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLVX ALIX-GFP S718/721A
FEATURES             Location/Qualifiers
     source          1..4200
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(2745..2747,2748..2750,2751..3464)
                     /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             join(2745..2747,2748..2750,2751..3461)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2790..2811)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3051..3070)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3398..3419
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        3478..3977
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3513..3536)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3910..3929
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(3998..4017)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gagctcgttt agtgaccgtc agatcgcctg gagaaggatc cgccaccatg atggcgacat
       61 tcatctcggt gcagctgaaa aagacctcag aggtggacct ggccaagccg ctggtgaagt
      121 tcatccagca gacttaccca agcggcgggg aagagcaggc ccagtactgc cgcgcggcgg
      181 aggagctcag caagctgcgc cgcgccgcag tcggtcgtcc gctggacaag cacgagggcg
      241 cgctcgagac gctcctgaga tattatgatc agatttgttc tattgaaccc aaattcccat
      301 tttctgaaaa tcagatctgc ttgacattta cctggaagga tgctttcgat aaaggttcac
      361 tttttggagg ctctgtaaaa ctggctcttg caagcttagg atatgaaaag agctgtgtgt
      421 tgttcaattg tgcagcctta gctagccaaa ttgcagcaga acagaactta gacaacgatg
      481 aaggattgaa aatcgctgct aaacattacc agtttgctag tggtgccttt ttacatatta
      541 aagagacggt tttatctgcc ttaagtcgag agccgaccgt ggacatatct ccagatactg
      601 ttgggaccct cagtcttatt atgctggcac aggctcaaga agtatttttt ttaaaagcca
      661 caagagataa aatgaaagat gccatcatag ctaaattggc taatcaggct gcagattatt
      721 ttggtgatgc tttcaaacag tgtcaataca aagatactct ccccaaggag gtgttccctg
      781 tcttggctgc aaagcactgt atcatgcagg ccaatgctga gtaccatcag tctatcctgg
      841 caaaacagca gaagaaattt ggagaagaaa ttgcaaggtt acagcatgca gcagaactga
      901 ttaaaacagt ggcatctcgc tatgatgaat atgttaatgt gaaggatttt tctgacaaaa
      961 tcaatcgtgc ccttgctgca gcaaagaagg ataatgactt catttatcat gatcgagttc
     1021 cagaccttaa agatctagat cctattggca aagccacact tgtgaaatct accccggtca
     1081 atgtacccat cagtcagaaa tttactgatc tgtttgagaa gatggttccc gtgtcagtac
     1141 agcagtcttt ggctgcctat aatcagagga aagccgattt ggttaacaga tcaattgctc
     1201 agatgagaga agccaccact ttggcaaatg gggtgctagc ttcccttaat cttccagcag
     1261 caattgaaga tgtgtctgga gacactgtac ctcagtctat attgactaaa tccagatctg
     1321 tgattgaaca gggaggcatc cagactgttg atcagttgat taaagaactg cctgaattac
     1381 tgcaacgaaa tagagaaatc ctagatgagt cattaaggtt gttggatgaa gaagaagcaa
     1441 ccgataatga tttaagagca aaatttaagg aacgttggca aaggacacca tccaatgaac
     1501 tgtataagcc tttaagagca gagggaacca acttcagaac agttttagat aaagctgtgc
     1561 aggcagatgg acaagtgaaa gaatgttacc agtctcatcg tgacaccatc gtgcttttgt
     1621 gtaagccaga gcctgagctg aatgctgcca tcccttctgc taatccagca aagaccatgc
     1681 agggcagtga ggttgtaaat gtcttaaaat ccttattgtc aaatcttgat gaagtaaaga
     1741 aggaaagaga gggtctggag aatgacttga aatctgtgaa ttttgacatg acaagcaagt
     1801 ttttgacagc cctggctcaa gatggtgtga taaatgaaga agctctttct gttactgaac
     1861 tagatcgagt ctatggaggt cttacaacta aagtccaaga atctctaaag aaacaggagg
     1921 gacttcttaa aaatattcag gtctcacatc aggaattttc aaaaatgaaa caatctaata
     1981 atgaagctaa cttaagagaa gaagttttga agaatttagc tactgcgtat gacaactttg
     2041 ttgaacttgt agctaatttg aaggaaggca caaagtttta caatgagttg actgaaatcc
     2101 tggtcaggtt ccagaacaaa tgcagtgata tagtttttgc acggaagaca gaaagagatg
     2161 aactcttaaa ggacttgcaa caaagcattg ccagagaacc tgctgctcct gctattccta
     2221 cacctgcgta tcagtcctca ccagcaggag gacatgcacc aactcctcca actccagcgc
     2281 caagaaccat gccgcctact aagccccagc ccccagccag gcctccacca cctgtgcttc
     2341 cagcaaatcg agctccttct gctactgctc catctccagt gggggctggg actgctgcgc
     2401 cagctccatc acaaacgcct ggctcagctc ctcctccaca ggcgcaggga ccaccctatc
     2461 ccacctatcc aggatatcct gggtattgcc aaatgcccat gcccatgggc tataatcctt
     2521 atgcgtatgg ccagtataat atgccatatc caccagtgta tcaccagagt cctggacagg
     2581 ctccataccc gggaccccag cagccttcat accccttccc tcagccccca cagcagtctt
     2641 actatccaca gcagcccggg gacgaggagc tcgagcgccg gcttcgcgag gaattggaac
     2701 ggcgccttcg cgaggagctg gaacgtcgac tccgcggctc ttccatggtg agcaagggcg
     2761 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2821 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2881 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2941 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3001 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3061 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3121 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3181 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3241 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3301 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3361 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3421 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaataagaa ttctaccggg
     3481 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     3541 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     3601 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     3661 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     3721 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     3781 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     3841 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     3901 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     3961 catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt
     4021 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4081 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4141 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
//
