LOCUS       sequence-110650-         900 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bicistronic expression of LifeAct-GFP fusion and neo3 resistance 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 900)
  TITLE     Piel Lab Plasmids
REFERENCE   2  (bases 1 to 900)
  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"
COMMENT     Sequence Label: pIRES LifeAct-GFP neo3
FEATURES             Location/Qualifiers
     source          1..900
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     52..71
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        52..70
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             122..172
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      188..197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(194..196,197..199,200..712)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(239..260)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(500..519)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     847..868
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcagagctct ctggctacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
       61 tcactatagg gagacccaag cttggtaccg agctcggatc gatatctgcg gcctagctag
      121 catgggcgtg gccgacctga tcaagaagtt cgagagcatc agcaaggagg agggcgaccc
      181 ccccgtggcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      241 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      301 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      361 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      421 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      481 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      541 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      601 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
      661 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      721 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      781 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
      841 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
//
