pAAV-CAG-PSmOrange3-P2A-EGFP

LOCUS      pAAV-CAG-PSmOrange3-P2A-EGFP             6289 bp DNA     circular SYN 20-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6289)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(941..1801)
                     /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(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    2448..2588
                     /label=ITR
     misc_feature    2458..2647
                     /label=ITR
     promoter        2942..3219
                     /label=chicken beta-actin promoter
     regulatory      3643..3652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3643..3652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3649..4356
                     /codon_start=1
                     /product="enhanced orange to far-red photoswitchable 
                     monomeric fluorescent protein (Subach et al., 2012)"
                     /label=PSmOrange3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGTVNGHEFEVEGKGEGHPYEG
                     FQTVKLKVTKGGPLPFAWDILSPLVTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEVRMRLKLKDGGHYTSEAKTTYKAKKSVLLPGAYIVGIKLDITSHNEDFTIVEQYERS
                     EARHSTGGKDELYK"
     CDS             4381..4437
                     /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             4450..5166
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    5191..5779
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5662..5673)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5822..5943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    6046..6235
                     /label=ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa ggcgcgcccc taggggagat ggccagccag cccattgacg
     2701 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2761 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2821 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2881 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2941 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     3001 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     3061 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
     3121 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     3181 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gacgctgcct
     3241 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     3301 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     3361 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     3421 gggccctttg tgcgggggga gcggctcggg gctgtccgcg gggggacggc tgccttcggg
     3481 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     3541 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     3601 tgctgtctca tcattttggc aaagaattgg ggatccggta ccgccaccat ggtgagcaag
     3661 ggcgaggaga ataatatggc catcattaag gagttcatgc gcttcaaggt gcacatggag
     3721 ggcactgtga acggccacga gttcgaggtc gagggcaagg gcgagggcca cccctacgag
     3781 ggctttcaga ccgttaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     3841 atcctgtccc ctctcgtcac ctacggctcc aaggcgtacg tgaagcaccc cgccgacatc
     3901 cccgactact tcaagctgtc cttccccgag ggattcaagt gggagcgtgt gatgaactac
     3961 gaggacggcg gcgtggtgac cgtgacccag gactcctctc tgcaggacgg cgagttcatc
     4021 tacaaggtga agatgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     4081 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     4141 gaggtcagga tgaggctgaa gctgaaggat ggcggccact acacctccga ggccaagact
     4201 acctacaagg ccaagaagtc cgtgctgctg cccggcgcct acatcgtcgg catcaagttg
     4261 gacattacct cacacaacga ggacttcacc atcgtggaac agtatgaacg cagcgaggcc
     4321 cgccactcca ccggcggcaa ggacgagctg tacaagcgac cggttcacgt gggaagtggg
     4381 gctacaaact tctcactgct gaaacaggcc ggggatgtgg aggaaaaccc aggaccacac
     4441 gtgactagtg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4501 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4561 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     4621 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     4681 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4741 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4801 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4861 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     4921 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     4981 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     5041 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     5101 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     5161 aagtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     5221 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5281 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5341 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5401 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5461 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5521 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     5581 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     5641 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5701 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     5761 ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgcttcg agcagacatg
     5821 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     5881 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     5941 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     6001 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     6061 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     6121 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     6181 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat
     6241 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgct
//