LOCUS       sequence-267744-        4182 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses fluorescent ALIX in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4182)
  TITLE     Plasmids for creating inducible cell lines expressing ALIX-GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4182)
  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            "Unpublished"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 WT
FEATURES             Location/Qualifiers
     source          1..4182
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(2742..2744,2745..2747,2748..3461)
                     /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(2742..2744,2745..2747,2748..3458)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2787..2808)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3048..3067)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3395..3416
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        3475..3974
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3510..3533)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3907..3926
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(3995..4014)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 ctcgtttagt gaccgtcaga tcgcctggag aaggatccgc caccatgatg gcgacattca
       61 tctcggtgca gctgaaaaag acctcagagg tggacctggc caagccgctg gtgaagttca
      121 tccagcagac ttacccaagc ggcggggaag agcaggccca gtactgccgc gcggcggagg
      181 agctcagcaa gctgcgccgc gccgcagtcg gtcgtccgct ggacaagcac gagggcgcgc
      241 tcgagacgct cctgagatat tatgatcaga tttgttctat tgaacccaaa ttcccatttt
      301 ctgaaaatca gatctgcttg acatttacct ggaaggatgc tttcgataaa ggttcacttt
      361 ttggaggctc tgtaaaactg gctcttgcaa gcttaggata tgaaaagagc tgtgtgttgt
      421 tcaattgtgc agccttagct agccaaattg cagcagaaca gaacttagac aacgatgaag
      481 gattgaaaat cgctgctaaa cattaccagt ttgctagtgg tgccttttta catattaaag
      541 agacggtttt atctgcctta agtcgagagc cgaccgtgga catatctcca gatactgttg
      601 ggaccctcag tcttattatg ctggcacagg ctcaagaagt atttttttta aaagccacaa
      661 gagataaaat gaaagatgcc atcatagcta aattggctaa tcaggctgca gattattttg
      721 gtgatgcttt caaacagtgt caatacaaag atactctccc caaggaggtg ttccctgtct
      781 tggctgcaaa gcactgtatc atgcaggcca atgctgagta ccatcagtct atcctggcaa
      841 aacagcagaa gaaatttgga gaagaaattg caaggttaca gcatgcagca gaactgatta
      901 aaacagtggc atctcgctat gatgaatatg ttaatgtgaa ggatttttct gacaaaatca
      961 atcgtgccct tgctgcagca aagaaggata atgacttcat ttatcatgat cgagttccag
     1021 accttaaaga tctagatcct attggcaaag ccacacttgt gaaatctacc ccggtcaatg
     1081 tacccatcag tcagaaattt actgatctgt ttgagaagat ggttcccgtg tcagtacagc
     1141 agtctttggc tgcctataat cagaggaaag ccgatttggt taacagatca attgctcaga
     1201 tgagagaagc caccactttg gcaaatgggg tgctagcttc ccttaatctt ccagcagcaa
     1261 ttgaagatgt gtctggagac actgtacctc agtctatatt gactaaatcc agatctgtga
     1321 ttgaacaggg aggcatccag actgttgatc agttgattaa agaactgcct gaattactgc
     1381 aacgaaatag agaaatccta gatgagtcat taaggttgtt ggatgaagaa gaagcaaccg
     1441 ataatgattt aagagcaaaa tttaaggaac gttggcaaag gacaccatcc aatgaactgt
     1501 ataagccttt aagagcagag ggaaccaact tcagaacagt tttagataaa gctgtgcagg
     1561 cagatggaca agtgaaagaa tgttaccagt ctcatcgtga caccatcgtg cttttgtgta
     1621 agccagagcc tgagctgaat gctgccatcc cttctgctaa tccagcaaag accatgcagg
     1681 gcagtgaggt tgtaaatgtc ttaaaatcct tattgtcaaa tcttgatgaa gtaaagaagg
     1741 aaagagaggg tctggagaat gacttgaaat ctgtgaattt tgacatgaca agcaagtttt
     1801 tgacagccct ggctcaagat ggtgtgataa atgaagaagc tctttctgtt actgaactag
     1861 atcgagtcta tggaggtctt acaactaaag tccaagaatc tctaaagaaa caggagggac
     1921 ttcttaaaaa tattcaggtc tcacatcagg aattttcaaa aatgaaacaa tctaataatg
     1981 aagctaactt aagagaagaa gttttgaaga atttagctac tgcgtatgac aactttgttg
     2041 aacttgtagc taatttgaag gaaggcacaa agttttacaa tgagttgact gaaatcctgg
     2101 tcaggttcca gaacaaatgc agtgatatag tttttgcacg gaagacagaa agagatgaac
     2161 tcttaaagga cttgcaacaa agcattgcca gagaacctag tgctccttca attcctacac
     2221 ctgcgtatca gtcctcacca gcaggaggac atgcaccaac tcctccaact ccagcgccaa
     2281 gaaccatgcc gcctactaag ccccagcccc cagccaggcc tccaccacct gtgcttccag
     2341 caaatcgagc tccttctgct actgctccat ctccagtggg ggctgggact gctgcgccag
     2401 ctccatcaca aacgcctggc tcagctcctc ctccacaggc gcagggacca ccctatccca
     2461 cctatccagg atatcctggg tattgccaaa tgcccatgcc catgggctat aatccttatg
     2521 cgtatggcca gtataatatg ccatatccac cagtgtatca ccagagtcct ggacaggctc
     2581 catacccggg accccagcag ccttcatacc ccttccctca gcccccacag cagtcttact
     2641 atccacagca gcccggggac gaggagctcg agcgccggct tcgcgaggaa ttggaacggc
     2701 gccttcgcga ggagctggaa cgtcgactcc gcggctcttc catggtgagc aagggcgagg
     2761 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2821 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2881 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2941 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3001 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3061 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3121 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3181 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3241 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3301 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3361 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3421 ccgccgggat cactctcggc atggacgagc tgtacaagta ataagaattc taccgggtag
     3481 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     3541 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     3601 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     3661 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     3721 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     3781 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     3841 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     3901 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     3961 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     4021 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4081 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4141 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aa
//
