LOCUS       sequence-354730-        6160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mCherry with microtubule-targeting sequence in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6160)
  AUTHORS   Kashiwagi S, Fujioka Y, Satoh AO, Yoshida A, Fujioka M, Nepal P, 
            Tsuzuki A, Aoki O, Paudel S, Sasajima H, Ohba Y
  TITLE     Folding Latency of Fluorescent Proteins Affects the Mitochondrial 
            Localization of Fusion Proteins.
  JOURNAL   Cell Struct Funct. 2019 Dec 26;44(2):183-194. doi: 
            10.1247/csf.19028. Epub 2019 Nov 15.
  PUBMED    31735740
REFERENCE   2  (bases 1 to 6160)
  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"; doi: "10.1247/csf.19028";
            journalName: "Cell Struct Funct"; date: "2019-12-26- 26"; volume: 
            "44"; issue: "2"; pages: "183-194"
COMMENT     Sequence Label: pFX-beta-tubulin-mCherry
FEATURES             Location/Qualifiers
     source          1..6160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1998..2705
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2143..2161)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2411..2430
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2948..3069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2985..3004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3039..3058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3076..3531)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3213..3234)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3425..3444
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3558..3662
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3664..4021
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3664..3855
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3687..3707)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3872..4007
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4056..4850
                     /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             4056..4850
                     /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(4110..4129)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4720..4739
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5038..5057)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5082..5129
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5458..6046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5947..5966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggttctag agccatgctc gagaccacca tgagggaaat cgtgcacttg caggccgggc
      661 agtgcggcaa ccaaatcggc gccaagtttt gggaggtgat cagcgatgag cacggcatcg
      721 accccacggg cacctaccac ggggacagcg acctgcagct ggaacgcatc aacgtgtact
      781 acaatgaggc caccggcggc aagtacgtgc cccgcgccgt gctcgtggat ctggagcccg
      841 gcaccatgga ctccgtgcgc tcggggccct tcgggcagat cttccggccg gacaacttcg
      901 ttttcggtca gagtggtgct gggaacaact gggccaaggg gcactacaca gaaggcgcgg
      961 agctggtgga ctcggtgctg gatgttgtga gaaaggaggc tgagagctgt gactgcctgc
     1021 agggtttcca gctgacccac tccctgggtg gggggactgg gtctgggatg ggtaccctcc
     1081 tcatcagcaa gatccgggag gagtacccag acaggatcat gaacacgttt agtgtggtgc
     1141 cttcgcccaa agtgtcagac acagtggtgg agccctacaa cgccaccctc tcagtccacc
     1201 agctcgtaga aaacacagac gagacctact gcattgataa cgaagctctc tacgacattt
     1261 gcttcagaac cctaaagctg accacgccca cctatggtga cctgaaccac ctggtgtctg
     1321 ctaccatgag tggggtcacc acctgcctgc gcttcccagg ccagctcaat gctgacctgc
     1381 ggaagctggc tgtgaacatg gtcccgtttc cccggctgca cttcttcatg cccggctttg
     1441 ccccactgac cagccggggc agccagcagt accgggcgct gaccgtgccc gagctcaccc
     1501 agcagatgtt tgatgccaag aacatgatgg ctgcctgcga cccccgccat ggccgctacc
     1561 tgacggttgc cgccgtgttc aggggccgca tgtccatgaa ggaggtggat gagcaaatgc
     1621 ttaatgtcca aaacaaaaac agcagctatt ttgttgagtg gatccccaac aatgtgaaaa
     1681 cggctgtctg tgacatccca cctcgggggc taaaaatgtc cgccaccttc attggcaaca
     1741 gcacggccat ccaggagctg ttcaagcgca tctccgagca gttcacggcc atgttccggc
     1801 gcaaggcctt cctgcactgg tacacgggcg agggcatgga cgagatggag ttcaccgagg
     1861 ccgagagcaa catgaatgac ctggtgtccg agtaccagca gtaccaggat gccacagccg
     1921 aggaggaggg cgagttcgag gaggaggctg aggaggaggt ggcctgcggc cgcggaggat
     1981 ccggaggaga attcaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     2041 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     2101 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2161 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2221 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     2281 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     2341 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     2401 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     2461 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     2521 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2581 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     2641 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2701 acaagggagg ctccggagga agatctctga ctgactgaga tgcgcggatg tcgaacggca
     2761 gttcggccga ataggccgac agccggccgt cggccgacga cacggccagg gccggcgggg
     2821 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2881 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2941 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3001 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3061 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3121 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3181 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3241 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3301 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3361 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3421 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3481 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3541 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3601 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3661 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3721 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3781 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3841 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3901 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3961 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4021 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4081 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4141 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4201 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4261 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4321 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4381 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4441 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4501 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4561 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4621 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4681 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4741 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4801 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4861 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4921 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4981 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5041 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5101 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5161 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5221 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5281 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5341 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5401 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5461 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5521 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5581 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5641 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5701 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5761 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5821 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5881 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5941 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6001 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6061 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6121 atcccctgat tctgtggata accgtattac cgccatgcat
//
