LOCUS       sequence-370678-        5436 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 5436)
  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 5436)
  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..5436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        418..692
                     /label=chicken beta-actin promoter
     primer_bind     971..993
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1055..1074
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1107..1116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1158..1179)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    1857..2445
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1910..1930)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2328..2339)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2488..2609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2499..2518)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2553..2572
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2771..2900
                     /label=AAV2 ITR
     rep_origin      complement(3137..3725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3217..3236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3896..4687,4688..4756))
                     /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     4519..4538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4757..4861)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4887..5342)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5024..5045)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5236..5255
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5414..5436,1..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 ggggggcgcg
      541 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
      601 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
      661 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
      721 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
      781 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat
      841 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
      901 tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
      961 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     1021 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     1081 catcattttg gcaaagaatt ggatccgcca ccatggtgag caagggcgag gagctgttca
     1141 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1201 tccgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ctcatctcca
     1261 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttaggc tacggcctga
     1321 tggtgttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1381 ccgaaggcta cgtccaggag cgcaccatct ctttcgagga cgacggttac tacaagaccc
     1441 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgtgctg aagggcatcg
     1501 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca
     1561 acgtctatat cacggccgac aagcagaaga acggcatcaa ggctaacttc aagatccgcc
     1621 acaacgttga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1681 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gtcctgagca
     1741 aagaccccaa cgagaagcgc gatcacatgg tcctgaagga gagggtgacc gccgccggga
     1801 ttacacatga catgaacgag ctgtacaagt aagaattcga tatcaagctt atcgataatc
     1861 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1921 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1981 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2041 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2101 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2161 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2221 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     2281 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2341 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2401 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     2461 ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2521 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2581 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2641 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2701 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     2761 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2821 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2881 gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2941 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3001 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3061 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3301 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3541 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3601 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3661 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3721 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3781 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4741 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4801 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4861 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     4921 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4981 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5041 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5101 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5161 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5221 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5281 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5341 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5401 gctattacgc cagctgcgcg ctcgctcgct cactga
//
