LOCUS       sequence-110763-         965 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bicistronic expression of LifeAct-GFP fusion and puro3 resistance 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 965)
  TITLE     Piel Lab Plasmids
REFERENCE   2  (bases 1 to 965)
  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 puro3
FEATURES             Location/Qualifiers
     source          1..965
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     51..70
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        51..69
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             121..171
                     /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      187..196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(193..195,196..198,199..711)
                     /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(238..259)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(499..518)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     845..866
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcagagctct tggctactag agaacccact gcttactggc ttatcgaaat taatacgact
       61 cactataggg agacccaagc ttggtaccga gctcggatcg atatctgcgg cctagctagc
      121 atgggcgtgg ccgacctgat caagaagttc gagagcatca gcaaggagga gggcgacccc
      181 cccgtggcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      241 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      301 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      361 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      421 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      481 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      541 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      601 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
      661 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
      721 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      781 cccgacaacc actacctgag cacccagtcc gccctgagca agaccccaac gagaagcgcg
      841 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
      901 tgtacaagta aagcggccgc atagataact gatccagtgt gctggaatta attcgctgtc
      961 tgcga
//
