pAAV-CAG-smURFP-P2A-GFP

LOCUS      pAAV-CAG-smURFP-P2A-GFP              5932 bp DNA     circular SYN 27-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5932)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Jun 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        279..556
                     /label=chicken beta-actin promoter
     regulatory      977..982
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             983..1381
                     /codon_start=1
                     /product="dimeric small ultra-red fluorescent protein 
                     derived from a cyanobacterial phycobiliprotein (Rodriguez 
                     et al., 2016)"
                     /label=smURFP
                     /translation="MAKTSEQRVNIATLLTENKKKIVDKASQDLWRRHPDLIAPGGIAF
                     SQRDRALCLRDYGWFLHLITFCLLAGDKGPIESIGLISIREMYNSLGVPVPAMMESIRC
                     LKEASLSLLDEEDANETAPYFDYIIKAMS"
     primer_bind     complement(1378..1404)
                     /label=0117-miRFP67-R
     CDS             1406..1462
                     /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             1475..2191
                     /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     complement(1546..1564)
                     /label=EGFP-R
     misc_feature    2216..2804
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2687..2698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2847..2968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3496..4084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4255..5115)
                     /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(5116..5220)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5246..5701)
                     /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 aattggatcc ggtaccgcca ccatggctaa gacttccgaa cagagggtga acattgctac
     1021 actgctgaca gaaaataaga agaaaatcgt ggataaggct tcccaggatc tgtggcggag
     1081 acacccagac ctgatcgcac caggaggaat tgctttctct cagagggacc gcgctctgtg
     1141 cctgcgagat tacggctggt tcctgcatct gatcaccttt tgtctgctgg ccggagataa
     1201 gggccccatc gagtctattg ggctgatcag tattcgagaa atgtataact cactgggagt
     1261 gcccgtccct gcaatgatgg agagcattag atgcctgaaa gaagccagcc tgtccctgct
     1321 ggacgaagag gacgccaacg agaccgcacc ctactttgat tacattatta aggctatgag
     1381 ccgaccggtt cacgtgggaa gtggggctac aaacttctca ctgctgaaac aggccgggga
     1441 tgtggaggaa aacccaggac cacacgtgac tagtgtgagc aagggcgagg agctgttcac
     1501 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1561 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1621 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     1681 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1741 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1801 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1861 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1921 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1981 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2041 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     2101 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2161 cactctcggc atggacgagc tgtacaagta agaattcgat atcaagctta tcgataatca
     2221 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2281 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2341 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2401 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2461 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2521 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2581 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     2641 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2701 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2761 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     2821 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2881 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2941 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3001 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     3061 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
     3121 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3181 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3241 tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3301 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3361 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3661 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3901 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4081 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4141 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4201 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4261 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4321 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4381 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4441 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4501 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4561 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4621 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4681 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4741 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4801 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4861 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4921 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4981 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5041 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5101 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5161 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5221 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5281 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5341 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5401 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5461 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5521 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5581 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5641 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5701 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5761 ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
     5821 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     5881 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta ct
//