LOCUS       sequence-207287-        6259 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6259)
  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 6259)
  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..6259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        223..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        528..805
                     /label=chicken beta-actin promoter
     intron          807..1824
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1748..1770
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1832..1851
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1934..2644
                     /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    2663..3251
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2716..2736)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3134..3145)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3286..3303)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3292..3499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3522..3662
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3522..3651
                     /label=AAV2 ITR
     rep_origin      3737..4192
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3824..3843)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4034..4055
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4209..4228)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4328..4350
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4388..4406)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4474..4578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4579..4647,4648..5439)
                     /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(4797..4816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5610..6198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6099..6118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgaaacaat tctgcaggaa tctagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg
      661 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
      721 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      781 taaaaagcga agcgcgcggc gggcggggag tcgctgcgac gctgccttcg ccccgtgccc
      841 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
      901 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
      961 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
     1021 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
     1081 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     1141 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
     1201 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     1261 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
     1321 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     1381 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
     1441 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
     1501 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
     1561 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
     1621 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
     1681 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     1741 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     1801 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     1861 cattttggca aagaattgat ttgataccgc gggccccgcg tggacggtac ccttcaggtc
     1921 gacgtcgggt ggcatggtgt ctaagggcga agagctgatc aaggaaaata tgcgtatgaa
     1981 ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa tgcacaggtg aaggagaagg
     2041 cagaccgtac gagggagtgc aaaccatgag gatcaaagtc atcgagggag gacccctgcc
     2101 atttgccttt gacattcttg ccacgtcgtt catgtatggc agccgtacct ttatcaagta
     2161 cccggccgac atccctgatt tctttaaaca gtcctttcct gagggtttta cttgggaaag
     2221 agttacgaga tacgaagatg gtggagtcgt caccgtcacg caggacacca gccttgagga
     2281 tggcgagctc gtctacaacg tcaaggtcag aggggtaaac tttccctcca atggtcccgt
     2341 gatgcagaag aagaccaagg gttgggagcc taatacagag atgatgtatc cagcagatgg
     2401 tggtctgaga ggatacactg acatcgcact gaaagttgat ggtggtggcc atctgcactg
     2461 caacttcgtg acaacttaca ggtcaaaaaa gaccgtcggg aacatcaaga tgcccggtgt
     2521 ccatgccgtt gatcaccgcc tggaaaggat cgaggagagt gacaatgaaa cctacgtagt
     2581 gcaaagagaa gtggcagttg ccaaatacag caaccttggt ggtggcatgg acgagctgta
     2641 caagtaatag aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2701 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2761 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2821 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2881 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2941 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3001 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3061 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     3121 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3181 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3241 cgcctccccg catcgatacc gagcgctgct cgagagatcg atctgcctcg actgtgcctt
     3301 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3361 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3421 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3481 atagcaggca tgctggggac acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3541 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3601 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3661 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3721 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3781 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     3841 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3901 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3961 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4021 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     4081 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     4141 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4201 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4261 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4321 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4381 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4441 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4501 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4561 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4621 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4681 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4741 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4801 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4861 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4921 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4981 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5041 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5101 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5161 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5221 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5281 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5341 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5401 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5461 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5521 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5581 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5641 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5701 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5761 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5821 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5881 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5941 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6001 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6061 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6121 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6181 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6241 tggccttttg ctcacatgt
//
