LOCUS       sequence-459871-        6159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Cre-dependent reporter mRuby2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6159)
  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 6159)
  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-DIO-mRuby2
FEATURES             Location/Qualifiers
     source          1..6159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..583
                     /label=chicken beta-actin promoter
     primer_bind     913..935
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     997..1016
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1083..1116)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1167..1200)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1212..1922)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     regulatory      1919..1928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1935..1968
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2019..2052
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2077..2665
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2130..2150)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2548..2559)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2697..3173
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2896..2915)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3213..3353
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3213..3342
                     /label=AAV2 ITR
     rep_origin      3428..3883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3515..3534)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3725..3746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3900..3919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4019..4041
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4079..4097)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4165..4269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4270..4338,4339..5130)
                     /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(4488..4507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5301..5889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5790..5809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5951..6080
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      421 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
      481 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      541 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      601 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      661 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      721 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      781 ggagcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
      841 cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac
      901 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
      961 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1021 ctcatcattt tggcaaagaa ttgcggccca acggtaccgg atcctctaga gtcgactccg
     1081 gaataacttc gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc
     1141 tgctattagc aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat
     1201 ggcgcgcctc atttgtacag ctcatccatt cctccgccaa ggccagcaaa cttggcaact
     1261 gcatgttccc gttgtaccac aaacatctcg ttgtcagact cctccagtcg ttccaagcgg
     1321 tgatcaaccg catgtatgcc aggcattttg atgttgccga cagttttctt gctcctgtac
     1381 gtggtcacaa atgagcatga caagtggcct cctccgtcaa ccttcagagc catgtgcgta
     1441 tagcctctca gtcctccatc tgcaggatac atcatctctg tgtttggctc ccaccctttg
     1501 gttttcttct gcatcacagg cccattggag ggaaagttca cccctctcac ttgcacgtga
     1561 tagaccagac atccgtcctc cagactagtg tcctgcatga ctgtgaccac accaccatcc
     1621 tcgtaacggg tcaccctttc ccatgtgaag ccctctggaa aggactgttt gaagaagtct
     1681 ggaatgcctt tggggtactt gatgaaggtc cgactgccat acatgaagct agtggcaaga
     1741 atgtcgaagg caaacgggag agggccacct tcaataacct tgatcctcat ggtctgagta
     1801 cccatgtagg gattcccttc accttcccct gtgcacttga actgatgccc attcacagac
     1861 ccttccatga ctaccttcat gcgcatgttc tccttgatga gttcctctcc cttgctcacc
     1921 atggtggcgc tagcataact tcgtataaag tatcctatac gaagttattt gccttaaccc
     1981 agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct
     2041 atacgaagtt atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     2101 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2161 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2221 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2281 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2341 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2401 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2461 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     2521 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2581 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2641 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     2701 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     2761 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     2821 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2881 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     2941 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3001 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3061 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3121 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3181 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3241 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3301 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3361 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3421 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3481 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3541 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3601 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3661 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3721 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3781 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3841 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3901 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3961 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4021 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4081 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4141 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4201 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4261 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4321 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4381 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4441 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4501 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4561 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4621 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4681 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4741 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4801 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4861 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4921 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4981 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5041 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5101 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5161 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5221 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5941 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
     6001 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     6061 ctccatcact aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat
     6121 tacggggtca ttagttcata gcccatatat ggagttccg
//
