LOCUS       sequence-383310-        3926 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the protein of emiRFP670-P2A-GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3926)
  AUTHORS   Zhang H, Papadaki S, Sun X, Wang X, Drobizhev M, Yao L, Rehbock M, 
            Koster RW, Wu L, Namikawa K, Piatkevich KD
  TITLE     Quantitative assessment of near-infrared fluorescent proteins.
  JOURNAL   Nat Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z.
  PUBMED    37666982
REFERENCE   2  (bases 1 to 3926)
  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: "Nat 
            Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z."
COMMENT     Sequence Label: pAAV-CAG-emiRFP670-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..3926
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        495..772
                     /label=chicken beta-actin promoter
     primer_bind     1051..1073
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1135..1154
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1193..1202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2153..2209
                     /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(2222..2224,2225..2938)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2222..2224,2225..2935)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2264..2285)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2525..2544)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2872..2893
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2963..3551
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3016..3036)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3434..3445)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3594..3715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3605..3624)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3659..3678
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cagtaccatg acgtcaggcg gccatcgtgg tacgccgatc cgtgccggtc gctgattatt
       61 atgcagctgc ccgctcgctc gctcactgag ccgccgggca aagcccgggc gtcgggcgac
      121 tttgttcgcc cgccctcagt gaccgaccga accgccccag agaggagtgc actcatcact
      181 agggtctgta gtaatgatta acccgccatg ctactatcta cgtagccatg ctctaggaag
      241 agtaccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      301 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      361 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      421 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      481 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      541 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
      601 gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc
      661 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      721 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      781 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      841 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      901 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
      961 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg
     1021 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1081 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1141 tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccggta ccgccaccat
     1201 ggccgagggc agcgtggcca gacagcccga cctgctgacc tgcgaacatg aagagatcca
     1261 cctcgccggc tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac atgatcatcg
     1321 cgtcatccag gccagcgcca acgccgcgga atttctgaat ctcggaagcg tactcggcgt
     1381 tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc tcgatcccac
     1441 cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc aatccctcta cggagtactg
     1501 cggtctgatg catcggcctc cggaaggcgg gctgatcatc gaactcgaac gtgccggccc
     1561 gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag cggatccgca cggcgggttc
     1621 actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg gctacgaccg
     1681 ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg gtattctccg agtgccatgt
     1741 gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg tcgactgtcc cgcagatggc
     1801 gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct atcagccggt
     1861 gccgctggag ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt cgggctgctt
     1921 cctgcgctcg atgtcgccgt gccatctgca gttcctgaag gacatgggcg tgcgcgccac
     1981 cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct gtcaccatta
     2041 tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg ccgaaaggat
     2101 cgcgacgcgg atcaccgcgc ttgagagccg accggttcac gtgggaagtg gggctacaaa
     2161 cttctcactg ctgaaacagg ccggggatgt ggaggaaaac ccaggaccac acgtgactag
     2221 tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2281 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2341 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2401 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2461 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2521 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2581 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2641 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2701 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2761 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2821 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2881 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga
     2941 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3001 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3061 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3121 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3181 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3241 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3301 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3361 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3421 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3481 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3541 cgcctccccg catcgatacc gtcgacccgg gcggccgctt cgagcagaca tgataagata
     3601 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3661 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3721 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     3781 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ctcccctaga gcatggctac
     3841 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     3901 gcatcctctc ttttgggcgg gtctcc
//
