LOCUS       sequence-478905-        6047 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  An AAV genome that ubiquitously expresses the fluorescent protein 
            mRuby2 from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6047)
  AUTHORS   Haziza S, Chrapkiewicz R, Zhang Y, Kruzhilin V, Li J, Li J, Delamare
            G, Swanson R, Buzsaki G, Kannan M, Vasan G, Lin MZ, Zeng H, Daigle 
            TL, Schnitzer MJ
  TITLE     Imaging high-frequency voltage dynamics in multiple neuron classes 
            of behaving mammals.
  JOURNAL   Cell, 188, 1-23, August 7, 2025. doi: 10.1016/j.cell.2025.06.028
  PUBMED    39185175
REFERENCE   2  (bases 1 to 6047)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell, 188, 
            1-23, August 7, 2025. doi: 10.1016/j.cell.2025.06.028"
COMMENT     Sequence Label: pAAV-CAG-mRuby2-WPRE
FEATURES             Location/Qualifiers
     source          1..6047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     992..1014
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1076..1095
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1129..1839
                     /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"
     misc_feature    1867..2455
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1920..1940)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2338..2349)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2482..2499)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2488..2712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(2922..3251)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2936..3071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3236..3256
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3497..4085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3577..3596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4256..5047,5048..5116))
                     /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     4879..4898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5117..5221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5289..5307
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5345..5367)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5467..5486
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5700..5719
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6009..6027
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6009..6026
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggtaccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
      481 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
      541 cggggggggg gggggggcgc gcgccggggg gggggggggg gggggggggg gggggsgggg
      601 sgrggsggrg aggkgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
      661 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
      721 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
      781 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
      841 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
      901 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac
      961 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     1021 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     1081 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tggatccgat ggtgtctaag
     1141 ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac
     1201 ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc cgtacatggg aactcaaacc
     1261 atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat tcttgccacg
     1321 tcgttcatgt atggcagccg tacttttatc aagtacccga aaggcattcc tgatttcttt
     1381 aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga agatggtgga
     1441 gtcgtcaccg tcatgcagga caccagcctt gaggatggct gtctcgttta ccacgtccaa
     1501 gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac caagggttgg
     1561 gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata cactcatatg
     1621 gcactgaaag ttgatggtgg tggccatctg tcttgctctt tcgtaacaac ttacaggtca
     1681 aaaaagaccg tcgggaacat caagatgccc ggtatccatg ccgttgatca ccgcctggaa
     1741 aggttagagg aaagtgacaa tgaaatgttc gtagtacaac gcgaacacgc agttgccaag
     1801 ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt aagaattcga tatcaagctt
     1861 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1921 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1981 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2041 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2101 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2161 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2221 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     2281 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2341 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2401 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     2461 taccgtcgac tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2521 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2581 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2641 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2701 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgactaga gcatggctac
     2761 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     2821 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2881 cgcccgggcg gcctcagtga gcgagcgagc gcgcagagct ttttgcaaaa gcctaggcct
     2941 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     3001 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     3061 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     3121 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     3181 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     3241 aactgacaca cattccacag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3301 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3661 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3901 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5221 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5281 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5341 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5401 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5461 actatgcggc atcagagcag attgtactga gagtgcacca ttcgacgctc tcccttatgc
     5521 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5581 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     5641 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     5701 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     5761 atgcgtccgg cgtagaggat ctggctagcg atgaccctgc tgattggttc gctgaccatt
     5821 tccgggtgcg ggacggcgtt accagaaact cagaaggttc gtccaaccaa accgactctg
     5881 acggcagttt acgagagaga tgatagggtc tgcttcagta agccagatgc tacacaatta
     5941 ggcttgtaca tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata
     6001 cacatacgat ttaggtgaca ctatagaata cacggaatta attctag
//
