LOCUS       sequence-236064-        4071 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CAG:gfp:P2A:mycNRNX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4071)
  TITLE     SYNPLA plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4071)
  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            "Unpublished"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: jk95
FEATURES             Location/Qualifiers
     source          1..4071
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        312..589
                     /label=chicken beta-actin promoter
     primer_bind     868..890
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     952..971
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1004..1013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1010..1012,1013..1015,1016..1726)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1010..1012,1013..1015,1016..1726)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1055..1076)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1316..1335)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1663..1684
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1736..1792
                     /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             1937..1966
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3158..3746
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3211..3231)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3629..3640)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3789..3910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3800..3819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3854..3873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcgt
       61 accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      121 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      181 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      241 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      301 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      361 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      421 gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag
      481 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
      541 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      601 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      661 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      721 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      781 gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg ggggacggct
      841 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
      901 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
      961 tggttattgt gctgtctcat cattttggca aagaattgga tctgccacca tggtgagcaa
     1021 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1081 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1141 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1201 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1261 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1321 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1381 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1441 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1501 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1561 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1621 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1681 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggaa gcggagctac
     1741 taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac ctatgtacca
     1801 gaggatgctc cggtgcggcg ccgatctggg atcgcccggg ggcggcagtg gcggcggcgc
     1861 aggggggcgc ctggccctga tctggatagt cccgctcacc ctcggcggcc tcctaggagt
     1921 ggcctggggg gaattcgaac aaaaactcat ctcagaagag gatctagcat ccagtttggg
     1981 agcgcaccac atccaccatt tccatggcag cagcaagcat cattcagtgc ctattgcaat
     2041 ctacaggtca ccagcatcct tgcgaggcgg acacgctggg acgacgtata tctttagcaa
     2101 aggtggtgga caaattacat ataagtggcc tcctaatgac cgacccagta cacgagcaga
     2161 caggctggcc atcgggttta gcactgttca gaaggaagca gtgttggtgc gtgtggacag
     2221 ttcctcaggc ctgggtgact accttgagct gcacatacac caaggaaaaa ttggagttaa
     2281 atttaatgtt gggacagatg acatcgccat tgaggagtcc aatgcaatca ttaatgatgg
     2341 aaaataccat gtagtgcgtt tcacgaggag tggtggcaat gccacgttac aggtggacag
     2401 ctggccagtc atcgaacgat accctgcagg gcgtcagctc acaatcttca atagccaagc
     2461 aaccataata attggcggga aagagcaggg ccagcccttc cagggccagc tctctgggct
     2521 ttactacaat ggcttgaaag ttctgaatat ggcggcagag aacgatgcca acatcgccat
     2581 agtgggaaat gtgaggctgg tcggtgaagt gccttcctct atgacaactg agtcgacagc
     2641 cactgccatg cagtccgaga tgtccacctc aatcatggag accaccacaa ccctggctac
     2701 cagcacagct cgacgaggaa agccccccac aaaggagcct atcagccaga ccacggatga
     2761 tatccttgtg gcctcggcag agtgtcccag cgacgatgag gacattgacc cctgtgagcc
     2821 gagctcaggt gggttagcca accccacccg agtgggcggc cgcgaaccat acccaggctc
     2881 ggcagaggtg atccgggagt ctagcagtac cactggcatg gtggtgggga ttgtcgcagc
     2941 agcagctctg tgcatcctca tcctcctcta tgccatgtac aagtacagga accgggatga
     3001 agggtcgtac cacgtggatg agagtcgaaa ctacatcagt aactcagcac agtccaatgg
     3061 ggctgtggtc aaggagatgc aacccagcag tgctaaaagt gccaacaaaa acaagaagaa
     3121 caaggataag gagtattatg tctagaagct tatcgataat caacctctgg attacaaaat
     3181 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3241 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3301 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3361 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3421 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3481 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3541 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     3601 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3661 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3721 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc
     3781 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3841 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3901 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3961 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     4021 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac a
//
