LOCUS       sequence-340997-        4596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRuby3 expressed under the CMV promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4596)
  AUTHORS   Zhong H, Ceballos CC, Massengill CI, Muniak MA, Ma L, Qin M, Petrie 
            SK, Mao T
  TITLE     High-fidelity, efficient, and reversible labeling of endogenous 
            proteins using CRISPR-based designer exon insertion.
  JOURNAL   Elife. 2021 Jun 8;10. pii: 64911. doi: 10.7554/eLife.64911.
  PUBMED    34100715
REFERENCE   2  (bases 1 to 4596)
  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: "Elife."; 
            date: "2021-06-8"; volume: "10. pii: 64911. doi"; pages: " 
            10.7554/eLife.64911"
COMMENT     Sequence Label: pKanCMV_mRuby3
FEATURES             Location/Qualifiers
     source          1..4596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    190..237
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     262..281
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(469..1263)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(469..1263)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(580..599)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1190..1209
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1298..1494)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(1306..1516)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1312..1447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1540..1560
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1585..1689)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1716..2171
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1803..1822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2013..2034
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2178..2299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2189..2208)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2243..2262
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2432..3142)
                     /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"
     regulatory      3139..3148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3187..3390)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3216..3236)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3391..3694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3869..4457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3949..3968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga
       61 gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt
      121 attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc
      181 cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca
      241 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt
      301 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
      361 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
      421 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
      481 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
      541 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
      601 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
      661 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
      721 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
      781 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
      841 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
      901 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
      961 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     1021 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     1081 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     1141 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     1201 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     1261 catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa
     1321 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     1381 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     1441 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     1501 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     1561 acagctggtt ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca
     1621 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1681 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     1741 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccaacg ccctagcgcc
     1801 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1861 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1921 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1981 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2041 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2101 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2161 gcttacaatt tacgccttaa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2221 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2281 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2341 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     2401 tgatcagtta tctagatccg gtggatccct acttgtacag ctcgtccatg ccaccaccaa
     2461 ggttgctgta tttggcaact gccacttctc tttgcactac gtaggtttca ttgtcactct
     2521 cctcgatcct ttccaggcgg tgatcaacgg catggacacc gggcatcttg atgttcccga
     2581 cggtcttttt tgacctgtaa gttgtcacga agttgcagtg cagatggcca ccaccatcaa
     2641 ctttcagtgc gatgtcagtg tatcctctca gaccaccatc tgctggatac atcatctctg
     2701 tattaggctc ccaacccttg gtcttcttct gcatcacggg accattggag ggaaagttta
     2761 cccctctgac cttgacgttg tagacgagct cgccatcctc aaggctggtg tcctgcgtga
     2821 cggtgacgac tccaccatct tcgtatctcg taactctttc ccaagtaaaa ccctcaggaa
     2881 aggactgttt aaagaaatca gggatgtcgg ccgggtactt gataaaggta cggctgccat
     2941 acatgaacga cgtggcaaga atgtcaaagg caaatggcag gggtcctccc tcgatgactt
     3001 tgatcctcat ggtttgcact ccctcgtacg gtctgccttc tccttcacct gtgcatttga
     3061 attggtggcc gttgaccgaa ccttccatga ccaccttcat acgcatattt tccttgatca
     3121 gctcttcgcc cttagacacc atggtggcga ccggtagcgc tagcggatct gacggttcac
     3181 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     3241 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     3301 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     3361 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     3421 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     3481 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3541 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3601 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3661 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3721 taatgacccc gtaattgatt actattaata actaatgcat ggcggtaata cggttatcca
     3781 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3841 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3901 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3961 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4021 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4081 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4141 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4201 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4261 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4321 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4381 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4441 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4501 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4561 tccttttaaa ttaaaaatga agttttaaat caatct
//
