LOCUS       sequence-338740-        6577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mRuby3 fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6577)
  AUTHORS   Sridharan S, Gajowa MA, Ogando MB, Jagadisan UK, Abdeladim L, 
            Sadahiro M, Bounds HA, Hendricks WD, Turney TS, Tayler I, Gopakumar 
            K, Oldenburg IA, Brohawn SG, Adesnik H
  TITLE     High-performance microbial opsins for spatially and temporally 
            precise perturbations of large neuronal networks.
  JOURNAL   Neuron. 2022 Jan 27. pii: S0896-6273(22)00008-3. doi: 
            10.1016/j.neuron.2022.01.008.
  PUBMED    35120626
REFERENCE   2  (bases 1 to 6577)
  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: "Neuron. 
            2022 Jan 27. pii: S0896-6273(22)00008-3. doi: 
            10.1016/j.neuron.2022.01.008."
COMMENT     Sequence Label: pCAG-mRuby3
FEATURES             Location/Qualifiers
     source          1..6577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1768..2478
                     /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"
     intron          2499..2631
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2614..2633
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2640..3228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2693..2713)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3111..3122)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3255..3272)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3261..3372
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3468..3488)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3468..3486)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3599..3618)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3664..3719
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3718..3737)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4080..4096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4080..4096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4093..4115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4104..4120
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4128..4134,4135..4152,4153..4158))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4173..4194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4253..4448
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4299..4434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4454..4588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4491..4510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4545..4564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4656..4673)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4827..5415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4907..4926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5586..6377,6378..6446))
                     /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     6209..6228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6447..6551)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
      421 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
      481 gatgggggcg gggggggggg gggsgcgcgc crggsggggg gggggggggg ggggggsggg
      541 gsggggsgrg gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc ggtaccgcgg
     1741 gcccgggatc caccggtagc ggccgctatg gtgtctaagg gcgaagagct gatcaaggaa
     1801 aatatgcgta tgaaggtggt catggaaggt tcggtcaacg gccaccaatt caaatgcaca
     1861 ggtgaaggag aaggcagacc gtacgaggga gtgcaaacca tgaggatcaa agtcatcgag
     1921 ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcatgta tggcagccgt
     1981 acctttatca agtacccggc cgacatccct gatttcttta aacagtcctt tcctgagggt
     2041 tttacttggg aaagagttac gagatacgaa gatggtggag tcgtcaccgt cacgcaggac
     2101 accagccttg aggatggcga gctcgtctac aacgtcaagg tcagaggggt aaactttccc
     2161 tccaatggtc ccgtgatgca gaagaagacc aagggttggg agcctaatac agagatgatg
     2221 tatccagcag atggtggtct gagaggatac actgacatcg cactgaaagt tgatggtggt
     2281 ggccatctgc actgcaactt cgtgacaact tacaggtcaa aaaagaccgt cgggaacatc
     2341 aagatgcccg gtgtccatgc cgttgatcac cgcctggaaa ggatcgagga gagtgacaat
     2401 gaaacctacg tagtgcaaag agaagtggca gttgccaaat acagcaacct tggtggtggc
     2461 atggacgagc tgtacaagtg aattcctcga ggccgcaggt aagtatcaag gttacaagac
     2521 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     2581 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtcgaca
     2641 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2701 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2761 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2821 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2881 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2941 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3001 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3061 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3121 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3181 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag
     3241 ctcggtacga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3301 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3361 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3421 ggacagcaag ggggaggatt gggaagacaa tagcccagct tttgttccct ttagtgaggg
     3481 ttaattgcgc gcttggcgta atcatkgtya tmssykwwty cwstcctcag gtgcaggctg
     3541 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     3601 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     3661 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     3721 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     3781 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     3841 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     3901 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     3961 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     4021 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     4081 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4141 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4201 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     4261 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4321 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4381 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4441 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4501 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4561 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     4621 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4681 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4741 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5641 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5701 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5761 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5821 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5881 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5941 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6001 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6061 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6121 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6181 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6241 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6301 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6361 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6421 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6481 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6541 ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
