LOCUS       sequence-207286-        5490 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Can be used to express mRuby3. Can also be used to create 
            adeno-associated virus for delivery of the mRuby3 sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5490)
  AUTHORS   Zhuang J, Wang Y, Ouellette ND, Turschak EE, Larsen RS, Takasaki KT,
            Daigle TL, Tasic B, Waters J, Zeng H, Reid RC
  TITLE     Laminar distribution and arbor density of two functional classes of 
            thalamic inputs to primary visual cortex.
  JOURNAL   Cell Rep. 2021 Oct 12;37(2):109826. doi: 
            10.1016/j.celrep.2021.109826.
  PUBMED    34644562
REFERENCE   2  (bases 1 to 5490)
  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: "Cell Rep.";
            date: "2021-10-12"; volume: "37(2):109826. doi"; pages: " 
            10.1016/j.celrep.2021.109826"
COMMENT     Sequence Label: AAV pCAG-mRuby3-WPRE
FEATURES             Location/Qualifiers
     source          1..5490
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     336..358
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     420..439
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             522..1232
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     misc_feature    1251..1839
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1304..1324)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1722..1733)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1874..1891)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1880..2087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2110..2250
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2110..2239
                     /label=AAV2 ITR
     rep_origin      2325..2780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2412..2431)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2622..2643
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2797..2816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2916..2938
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2976..2994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3062..3166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3167..3235,3236..4027)
                     /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(3385..3404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4198..4786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4687..4706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4848..4977
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        5070..5373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
       61 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
      121 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
      181 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
      241 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
      301 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
      361 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
      421 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgattt gataccgcgg
      481 gccccgcgtg gacggtaccc ttcaggtcga cgtcgggtgg catggtgtct aagggcgaag
      541 agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc aacggccacc
      601 aattcaaatg cacaggtgaa ggagaaggca gaccgtacga gggagtgcaa accatgagga
      661 tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc acgtcgttca
      721 tgtatggcag ccgtaccttt atcaagtacc cggccgacat ccctgatttc tttaaacagt
      781 cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt ggagtcgtca
      841 ccgtcacgca ggacaccagc cttgaggatg gcgagctcgt ctacaacgtc aaggtcagag
      901 gggtaaactt tccctccaat ggtcccgtga tgcagaagaa gaccaagggt tgggagccta
      961 atacagagat gatgtatcca gcagatggtg gtctgagagg atacactgac atcgcactga
     1021 aagttgatgg tggtggccat ctgcactgca acttcgtgac aacttacagg tcaaaaaaga
     1081 ccgtcgggaa catcaagatg cccggtgtcc atgccgttga tcaccgcctg gaaaggatcg
     1141 aggagagtga caatgaaacc tacgtagtgc aaagagaagt ggcagttgcc aaatacagca
     1201 accttggtgg tggcatggac gagctgtaca agtaatagaa gcttatcgat aatcaacctc
     1261 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1321 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1381 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     1441 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     1501 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     1561 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     1621 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     1681 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     1741 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     1801 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     1861 agagatcgat ctgcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1921 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1981 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2041 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggacac gtgcggaccg
     2101 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2161 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2221 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     2281 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     2341 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2401 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2461 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2521 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2581 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2641 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2701 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2761 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     2821 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2881 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2941 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     3001 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3061 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3121 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3181 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3241 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3301 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3361 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3421 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3481 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3541 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3601 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3661 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3721 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3781 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3841 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3901 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3961 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4021 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4081 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4141 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4201 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4261 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4321 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4381 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4441 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4501 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4561 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4621 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4681 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4741 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4801 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
     4861 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
     4921 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     4981 gccgcacgcg aaacaattct gcaggaatct agttattaat agtaatcaat tacggggtca
     5041 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     5101 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5161 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     5221 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     5281 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     5341 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     5401 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     5461 ttttgtgcag cgatgggggc gggggggggg
//
