LOCUS       sequence-207886-        5787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of N-terminal fusion of mRuby3 to actin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  AUTHORS   Bajar BT, Wang ES, Lam AJ, Kim BB, Jacobs CL, Howe ES, Davidson MW, 
            Lin MZ, Chu J
  TITLE     Improving brightness and photostability of green and red fluorescent
            proteins for live cell imaging and FRET reporting.
  JOURNAL   Sci Rep. 2016 Feb 16;6:20889. doi: 10.1038/srep20889.
  PUBMED    26879144
REFERENCE   2  (bases 1 to 5787)
  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: "Sci Rep."; 
            date: "2016-02-16"; volume: "6:20889. doi"; pages: " 
            10.1038/srep20889"
COMMENT     Sequence Label: pKanCMV-mRuby3-18aa-actin
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        224..527
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        528..731
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     682..702
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      770..779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             776..1486
                     /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"
     polyA_signal    2795..2916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2832..2851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2886..2905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2923..3378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3060..3081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3272..3291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3405..3509
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3511..3868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3534..3554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3719..3854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3903..4697
                     /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             3903..4697
                     /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(3957..3976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4567..4586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4885..4904)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4929..4976
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5305..5787,1..106)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
       61 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttmt twmywgtrat
      181 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      241 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      301 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      361 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      421 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      481 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      541 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      601 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      661 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      721 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
      781 gtctaagggc gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc
      841 ggtcaacggc caccaattca aatgcacagg tgaaggagaa ggcagaccgt acgagggagt
      901 gcaaaccatg aggatcaaag tcatcgaggg aggacccctg ccatttgcct ttgacattct
      961 tgccacgtcg ttcatgtatg gcagccgtac ctttatcaag tacccggccg acatccctga
     1021 tttctttaaa cagtcctttc ctgagggttt tacttgggaa agagttacga gatacgaaga
     1081 tggtggagtc gtcaccgtca cgcaggacac cagccttgag gatggcgagc tcgtctacaa
     1141 cgtcaaggtc agaggggtaa actttccctc caatggtccc gtgatgcaga agaagaccaa
     1201 gggttgggag cctaatacag agatgatgta tccagcagat ggtggtctga gaggatacac
     1261 tgacatcgca ctgaaagttg atggtggtgg ccatctgcac tgcaacttcg tgacaactta
     1321 caggtcaaaa aagaccgtcg ggaacatcaa gatgcccggt gtccatgccg ttgatcaccg
     1381 cctggaaagg atcgaggaga gtgacaatga aacctacgta gtgcaaagag aagtggcagt
     1441 tgccaaatac agcaaccttg gtggtggcat ggacgagctg tacaagtccg gactcagatc
     1501 tggcagcggt ggaggcagcg catccggcgg aagcggaagc gatgatgata tcgccgcgct
     1561 cgtcgtcgac aacggctccg gcatgtgcaa ggccggcttc gcgggcgacg atgccccccg
     1621 ggccgtcttc ccctccatcg tggggcgccc caggcaccag ggcgtgatgg tgggcatggg
     1681 tcagaaggat tcctatgtgg gcgacgaggc ccagagcaag agaggcatcc tcaccctgaa
     1741 gtaccccatc gagcacggca tcgtcaccaa ctgggacgac atggagaaaa tctggcacca
     1801 caccttctac aatgagctgc gtgtggctcc cgaggagcac cccgtgctgc tgaccgaggc
     1861 ccccctgaac cccaaggcca accgcgagaa gatgacccag atcatgtttg agaccttcaa
     1921 caccccagcc atgtacgttg ctatccaggc tgtgctatcc ctgtacgcct ctggccgtac
     1981 cactggcatc gtgatggact ccggtgacgg ggtcacccac actgtgccca tctacgaggg
     2041 gtatgccctc ccccatgcca tcctgcgtct ggacctggct ggccgggacc tgactgacta
     2101 cctcatgaag atcctcaccg agcgcggcta cagcttcacc accacggccg agcgggaaat
     2161 cgtgcgtgac attaaggaga agctgtgcta cgtcgccctg gacttcgagc aagagatggc
     2221 cacggctgct tccagctcct ccctggagaa gagctacgag ctgcctgacg gccaggtcat
     2281 caccattggc aatgagcggt tccgctgccc tgaggcactc ttccagcctt ccttcctggg
     2341 catggagtcc tgtggcatcc acgaaactac cttcaactcc atcatgaagt gtgacgtgga
     2401 catccgcaaa gacctgtacg ccaacacagt gctgtctggc ggcaccacca tgtaccctgg
     2461 cattgccgac aggatgcaga aggagatcac tgccctggca cccagcacaa tgaagatcaa
     2521 gatcattgct cctcctgagc gcaagtactc cgtgtggatc ggcggctcca tcctggcctc
     2581 gctgtccacc ttccagcaga tgtggatcag caagcaggag tatgacgagt ccggcccctc
     2641 catcgtccac cgcaaatgct tctagggatc caccggatct agataactga tcataatcag
     2701 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2761 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2821 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2881 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2941 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3001 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3061 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3121 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3181 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3241 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3301 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3361 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3481 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3541 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3601 gcatctcaat tagtcagcaa ccakrkkysg aaagtcccca ggctccccag caggcagaag
     3661 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3721 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3781 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3841 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3901 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3961 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4021 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4081 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4141 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4201 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4261 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4321 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4381 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4441 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4501 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4561 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4621 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4681 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4741 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4801 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4861 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4921 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4981 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5041 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5101 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5161 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5221 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5521 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5581 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5641 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5701 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5761 cggaacagga gagcgcacga gggagct
//
