pAAV-CAG-mfRFP-P2A-GFP

LOCUS       pAAV-CAG-mfRFP-P2A-GFP             6264 bp DNA     circular SYN 26-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Jul 6, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     961..1002
                     /label=2022.10.12 mCarmine F
     primer_bind     975..999
                     /label=0908-EB3-F
     regulatory      976..981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             982..1713
                     /codon_start=1
                     /label=mfRFP
                     /translation="MVSKGEELIKENMPLKLYMEGTVNNHHFKCTAEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATSFMYGSHTFIDHTPGIPDFWKQSFPEGFTWERVTTYEDGG
                     VLIATQDTSLHDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEAHTELLYPTDGGLEGRTA
                     LALKLVGGGHLLCNFKTIYRSKKPAKNLKMPGVYYVDYRLERIEEADNETYVEQHEVAV
                     ARYCDLPTKLGHKLKGMDELYK"
     primer_bind     complement(1692..1713)
                     /label=0908-EB3-R
     primer_bind     complement(1695..1736)
                     /label=2022.10.12 mCarmine R
     CDS             1738..1794
                     /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             1807..2523
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     1807..1821
                     /label=0908-EB3-F
     primer_bind     complement(2502..2521)
                     /label=2022.10.12 mCarmine R
     misc_feature    2548..3136
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3019..3030)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3179..3300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3828..4416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4587..5447)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5448..5552)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5578..6033)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tctacgtagc catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat
       61 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      121 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      181 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      241 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      301 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      361 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
      421 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
      481 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      541 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      601 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      661 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      721 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      781 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
      841 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
      901 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
      961 attggatccg gtaccgccac catggtgagc aagggcgagg agctgattaa ggagaacatg
     1021 ccactgaagc tgtacatgga gggcaccgtt aacaatcacc acttcaagtg cacagctgag
     1081 ggcgaaggca agccctacga gggcacccag acccagagaa tcaaggtggt cgagggcggc
     1141 cctctcccct tcgccttcga catcctggct accagtttca tgtacggcag tcacaccttc
     1201 atcgaccaca ccccgggcat ccccgacttc tggaagcagt ccttccccga gggcttcacg
     1261 tgggagagag tcaccacata cgaagacggg ggcgtcctga tcgcgaccca ggacaccagc
     1321 ctccatgacg gctgcctcat ctacaacgtc aagttgagag gggtgaactt cccatccaac
     1381 ggccctgtga tgcaaaagaa aacactcggc tgggaggcac acaccgagct tttgtacccc
     1441 actgacggcg gtctggaagg cagaaccgcg ctggccctga agcttgtggg cgggggccac
     1501 ctattgtgta acttcaagac tatatacaga tccaagaaac ccgctaagaa cctcaagatg
     1561 cccggcgtct actatgtgga ctacagactg gaaagaatcg aggaggccga caatgagacc
     1621 tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctac caaactgggg
     1681 cacaagctta agggcatgga cgagctgtac aagcgaccgg ttcacgtggg aagtggggct
     1741 acaaacttct cactgctgaa acaggccggg gatgtggagg aaaacccagg accacacgtg
     1801 actagtgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1861 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1921 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1981 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2041 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2101 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2161 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2221 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2281 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2341 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2401 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2461 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2521 taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     2581 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2641 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2701 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2761 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2821 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2881 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2941 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3001 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3061 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3121 tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc agacatgata
     3181 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3241 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3301 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     3361 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg
     3421 ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     3481 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3541 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa
     3601 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3661 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3721 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3781 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3841 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3901 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3961 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4021 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4081 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4141 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4201 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4261 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4321 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4381 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4441 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4501 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4561 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4621 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4681 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4741 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4801 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4861 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4921 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4981 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5041 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5101 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5161 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5221 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5281 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5341 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5401 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5461 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5521 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     5581 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5641 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5701 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5761 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5821 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5881 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5941 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6001 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     6061 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
     6121 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     6181 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
     6241 taatgattaa cccgccatgc tact
//