LOCUS       sequence-146553-         968 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid for expression of wild-type or mutant cDNA fragments 
            (mutation reporter) or complete cDNAs in frame with superfolderGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 968)
  TITLE     pCS2+MCS-P2A-sfGFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 968)
  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            "Unpublished"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: pCS2+MCS-P2A-sfGFP
FEATURES             Location/Qualifiers
     source          1..968
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        63..81
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     63..80
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             183..239
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(246..248,249..962)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             246..959
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(276..304)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     899..919
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 ctatataagc aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct
       61 tgatttaggt gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt
      121 cgaattcaag gcctctcgag cctctagatt aattaagcgc atgcggcgcg ccaggaagcg
      181 gagctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggacctg
      241 gatccatgag caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
      301 atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt gatgctacaa
      361 acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt ccgtggccaa
      421 cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga
      481 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat
      541 ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa ggtgataccc
      601 ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac attcttggac
      661 acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac aaacaaaaga
      721 atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc gttcaactag
      781 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
      841 attacctgtc gacacaatct gtcctgtcga aagatcccaa cgaaaagcgt gaccacatgg
      901 tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat
      961 aatctaga
//
