LOCUS       sequence-376988-        5438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive CAG-driven expression of hfYFP in animals using AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5438)
  AUTHORS   Campbell BC, Paez-Segala MG, Looger LL, Petsko GA, Liu CF
  TITLE     Chemically stable fluorescent proteins for advanced microscopy.
  JOURNAL   Nat Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7.
  PUBMED    36344833
REFERENCE   2  (bases 1 to 5438)
  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"; journalName: "Nat 
            Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7."
COMMENT     Sequence Label: pAAV-CAG-hfYFP
FEATURES             Location/Qualifiers
     source          1..5438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     973..995
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1057..1076
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1109..1118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1160..1181)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    1859..2447
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1912..1932)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2330..2341)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2490..2611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2501..2520)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2555..2574
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2773..2902
                     /label=AAV2 ITR
     rep_origin      complement(3139..3727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3219..3238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3898..4689,4690..4758))
                     /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     4521..4540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4759..4863)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4889..5344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5026..5047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5238..5257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5416..5438,1..107)
                     /label=AAV2 ITR
ORIGIN
        1 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
       61 gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt agttaatgat
      121 taacccgcca tgctacttat ctacgtagcc atgctctagg aagagtacca ttgacgtcaa
      181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
      421 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      481 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggggnn
      541 sgcgccrggs ggggsggggs ggggsgrggg gsggggsggg gsgrggsggr grggkgsggs
      601 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
      661 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
      721 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
      781 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
      841 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
      901 tttgtgcggg gggagcggct cggggctgtc cgcgggggga cggctgcctt cgggggggac
      961 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
     1021 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1081 ctcatcattt tggcaaagaa ttggatccgc caccatggtg agcaagggcg aggagctgtt
     1141 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1201 cgtccgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agctcatctc
     1261 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttag gctacggcct
     1321 gatggtgttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1381 gcccgaaggc tacgtccagg agcgcaccat ctctttcgag gacgacggtt actacaagac
     1441 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgtgc tgaagggcat
     1501 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca
     1561 caacgtctat atcacggccg acaagcagaa gaacggcatc aaggctaact tcaagatccg
     1621 ccacaacgtt gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     1681 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgtcctgag
     1741 caaagacccc aacgagaagc gcgatcacat ggtcctgaag gagagggtga ccgccgccgg
     1801 gattacacat gacatgaacg agctgtacaa gtaagaattc gatatcaagc ttatcgataa
     1861 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1921 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1981 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2041 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2101 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2161 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2221 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     2281 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2341 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2401 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     2461 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2521 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2581 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2641 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2701 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     2761 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2821 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2881 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2941 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3001 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3061 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3301 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3541 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3661 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3721 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3781 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3841 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3901 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3961 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4021 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4081 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4141 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4201 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4261 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4321 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4381 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4441 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4501 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4561 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4621 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4681 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4741 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4801 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4861 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4921 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4981 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5041 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5101 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5161 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5221 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5281 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5341 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5401 tcgctattac gccagctgcg cgctcgctcg ctcactga
//
