LOCUS       sequence-107173-         777 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AdEasy Shuttle vector modified for multicistronic expression. 
            Contains T2A, F2A and Venus tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 777)
  TITLE     Modified AdEasy shuttle vectors for multicistronic expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 777)
  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            "Unpublished"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: pShuttle-CMV-F2A-T2A-Venus
FEATURES             Location/Qualifiers
     source          1..777
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             57..122
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             153..206
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      213..222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(219..221,222..224,225..737)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(264..285)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(525..544)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gtgaaccgtc agatccgcta gagatctggt accgtcgacg cggccgcgga agcggagtga
       61 aacagacttt gaattttgac cttctcaagt tggcgggaga cgtggagtcc aaccctggac
      121 ctaggcctgg aagcggaacg cgtggaagcg gagagggcag aggaagtctg ctaacatgcg
      181 gtgacgtcga ggagaatcct ggacctctcg aggccaccat ggtgagcaag ggcgaggagc
      241 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      301 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
      361 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
      421 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      481 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      541 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      601 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      661 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
      721 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaaca
//
