LOCUS       sequence-201341-        5882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome that ubiquitously expresses the fluorescent protein 
            eYFP from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5882)
  AUTHORS   Challis RC, Ravindra Kumar S, Chan KY, Challis C, Beadle K, Jang MJ,
            Kim HM, Rajendran PS, Tompkins JD, Shivkumar K, Deverman BE, 
            Gradinaru V
  TITLE     Systemic AAV vectors for widespread and targeted gene delivery in 
            rodents.
  JOURNAL   Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3.
  PUBMED    30626963
REFERENCE   2  (bases 1 to 5882)
  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"; doi: "10"; journalName: 
            "Nat Protoc"; date: "2019-02"; volume: "14"; issue: "2"; pages: 
            "379-414"
COMMENT     Sequence Label: pAAV-CAG-eYFP
FEATURES             Location/Qualifiers
     source          1..5882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     431..453
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     515..534
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(577..579,580..582,583..1296)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(622..643)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(883..902)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1230..1251
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1321..1909
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1374..1394)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1792..1803)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1941..2417
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2140..2159)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2457..2597
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2457..2586
                     /label=AAV2 ITR
     rep_origin      2672..3127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2759..2778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2969..2990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3144..3163)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3263..3285
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3323..3341)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3409..3513
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3514..3582,3583..4374)
                     /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"
     primer_bind     complement(3732..3751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4545..5133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5034..5053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5195..5324
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        5404..5707
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5709..5882,1..101)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
       61 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      121 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      181 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
      241 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
      301 agcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
      361 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
      421 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
      481 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
      541 catcattttg gcaaagaatt gcggcccaac ggtaccatgg tgagcaaggg cgaggagctg
      601 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      661 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      721 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
      781 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      841 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      901 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      961 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1021 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1081 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1141 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     1201 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1261 gggatcactc tcggcatgga cgagctgtac aagtaagaat tcgatatcaa gcttatcgat
     1321 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1381 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     1441 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     1501 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     1561 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     1621 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1681 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     1741 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     1801 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     1861 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     1921 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     1981 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     2041 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     2101 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     2161 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     2221 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     2281 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     2341 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     2401 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     2461 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     2521 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     2581 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2641 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     2701 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     2761 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2821 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2881 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2941 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3001 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     3061 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3121 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3181 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3241 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3301 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3361 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3421 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3481 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3541 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3601 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3661 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3721 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3781 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3841 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3901 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3961 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4021 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4081 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4141 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4201 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4261 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4321 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4381 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4441 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4501 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4561 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4621 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4681 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4741 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4801 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4861 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4921 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4981 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5041 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5101 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5161 ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg
     5221 ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     5281 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgtaa
     5341 cactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     5401 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     5461 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     5521 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     5581 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     5641 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     5701 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     5761 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     5821 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
     5881 ga
//
