LOCUS       sequence-110666-         984 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bicistronic expression of LifeAct-mCherry fusion and Hygro 
            resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 984)
  TITLE     Piel Lab Plasmids
REFERENCE   2  (bases 1 to 984)
  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 Hygro3 LifeAct-mChe
FEATURES             Location/Qualifiers
     source          1..984
                     /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             129..179
                     /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      195..204
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             201..911
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(346..364)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     614..633
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 gcagagctct ctggctacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
       61 tcactatagg gagacccaag cttggtaccg agctcggatc tgtacaggcc ttaagccggc
      121 gcgctagcat gggcgtggcc gacctgatca agaagttcga gagcatcagc aaggaggagg
      181 gcgacccccc cgtggccacc atggtgagca agggcgagga ggataacatg gccatcatca
      241 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
      301 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
      361 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
      421 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
      481 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
      541 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
      601 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
      661 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
      721 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
      781 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
      841 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
      901 tgtacaagtg aggatccgat atcgtacgtt aacatagata actgatccag tgtgctggaa
      961 ttaattcgct gtctgcgagg gcca
//
