pAAV-CAG-BDFP1.8-P2A-GFP

LOCUS       pAAV-CAG-BDFP1.8-P2A-GFP            5989 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 5989)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Jun 23, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5989
                     /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..1438
                     /codon_start=1
                     /label=BDFP1.8
                     /translation="MANREVVETLKELLADGEKRVQVAGVIGTNAAEVVKTAVSLLFQE
                     YPELVSPGGCAYTTRRYNKCVRDMNYFLRMCSYAIVAGGASVLDGRMLAGLRDTFNSLG
                     IPLGPVARGIQLMKKIVKEKLATAGMTNIAFVDEPFDYIARVISETEI"
     CDS             1463..1519
                     /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             1532..2248
                     /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(1603..1621)
                     /label=EGFP-R
     misc_feature    2273..2861
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2744..2755)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2904..3025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3553..4141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4312..5172)
                     /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(5173..5277)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5303..5758)
                     /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 ccatggccaa tcgcgaggtt gttgagaccc tgaaggagct
     1021 cctggctgac ggggagaaga gagtgcaggt ggctggagtg attgggacta acgctgccga
     1081 agttgtcaag accgccgtgt cacttctgtt tcaggagtac ccagagttgg tgtcccctgg
     1141 cgggtgtgcc tatacaacaa ggcgctacaa caagtgtgtg agggacatga actacttcct
     1201 caggatgtgt agctacgcca tcgtggccgg gggcgccagc gtcttggatg gaaggatgct
     1261 ggccgggctg cgggatacat tcaactccct cggcatccct ctgggtcccg tggcccgggg
     1321 catccaactg atgaagaaga tcgtcaaaga gaaactggcg acagccggca tgactaacat
     1381 tgcatttgtc gatgagccct ttgactacat cgcccgtgtg attagtgaaa cggaaattcg
     1441 accggttcac gtgggaagtg gggctacaaa cttctcactg ctgaaacagg ccggggatgt
     1501 ggaggaaaac ccaggaccac acgtgactag tgtgagcaag ggcgaggagc tgttcaccgg
     1561 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1621 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1681 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     1741 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1801 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1861 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1921 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1981 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2041 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2101 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2161 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2221 tctcggcatg gacgagctgt acaagtaaga attcgatatc aagcttatcg ataatcaacc
     2281 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2341 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2401 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2461 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2521 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2581 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2641 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     2701 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2761 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2821 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacccgg
     2881 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2941 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3001 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3061 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     3121 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     3181 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3241 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3301 gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3361 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3421 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3481 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3541 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3601 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3661 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3721 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3781 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3841 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3901 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3961 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4021 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4081 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4141 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4201 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4261 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4321 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4381 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4441 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4501 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4561 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4621 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4681 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4741 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4801 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4861 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4921 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4981 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5041 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5101 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5161 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5221 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5281 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     5341 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5401 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5461 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5521 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5581 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5641 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5701 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     5761 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5821 ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     5881 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     5941 tccatcacta ggggttcctt gtagttaatg attaacccgc catgctact
//