LOCUS       sequence-354725-        6385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express iRFP with microtubule-targeting sequence in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6385)
  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 6385)
  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-iRFP
FEATURES             Location/Qualifiers
     source          1..6385
                     /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"
     polyA_signal    3173..3294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3210..3229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3264..3283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3301..3756)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3438..3459)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3650..3669
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3783..3887
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3889..4246
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3889..4080
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3912..3932)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4097..4232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4281..5075
                     /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             4281..5075
                     /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(4335..4354)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4945..4964
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5263..5282)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5307..5354
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5683..6271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6172..6191
                     /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 gccgaaggga gcgtggcaag acagcccgat ctgctgacat
     2041 gcgacgacga accaatccac attccaggag ctatccagcc tcatggactg ctgctggctc
     2101 tggccgctga catgactatc gtggcaggct ctgataacct gccagagctg accgggctgg
     2161 ctatcggagc actgattggc cggtccgcag ccgacgtgtt cgattctgag actcacaata
     2221 gactgaccat cgccctggct gaacctgggg ctgcagtggg agccccaatt accgtcggct
     2281 tcacaatgcg aaaggacgct ggctttatcg ggtcctggca ccgccatgat cagctgattt
     2341 ttctggagct ggaaccacct cagagagacg tggcagagcc acaggctttc tttcggagaa
     2401 caaacagcgc catcaggcgc ctgcaggccg ctgagactct ggaatccgct tgcgcagccg
     2461 ctgcacagga agtgaggaag atcaccgggt tcgatcgcgt catgatctac agattcgcct
     2521 cagactttag cggagaagtg atcgcagaag atcgctgtgc cgaggtcgaa agtaaactgg
     2581 gcctgcatta cccagcctca accgtgcctg cacaggcacg acggctgtat acaatcaacc
     2641 ccgtgcggat cattcccgac attaattaca gaccagtgcc cgtcacacct gacctgaacc
     2701 cagtgactgg gaggcccatc gatctgagct tcgccattct gcgcagcgtg tcccctgtcc
     2761 acctggagtt tatgaggaat atcggaatgc atggcaccat gtctatcagt attctgagag
     2821 gcgaaaggct gtgggggctg atcgtgtgcc accatcggac accttactat gtggacctgg
     2881 acggcaggca ggcttgtgaa ctggtggctc aggtgctggc ttggcagatt ggcgtgatgg
     2941 aagaaagatc tctgactgac tgagatgcgc ggatgtcgaa cggcagttcg gccgaatagg
     3001 ccgacagccg gccgtcggcc gacgacacgg ccagggccgg cggggatcca ccggatctag
     3061 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3121 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3181 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3241 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     3301 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3361 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3421 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3481 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3541 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3601 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3661 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3721 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3781 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3841 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3901 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3961 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4021 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4081 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4141 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4201 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4261 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4321 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4381 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4441 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4501 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4561 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4621 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4681 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4741 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4801 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4861 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4921 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4981 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5041 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5101 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5161 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5221 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5281 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5341 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5401 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5461 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5521 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5701 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5761 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5821 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5881 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5941 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6001 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6061 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6121 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6181 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6241 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6301 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6361 ggataaccgt attaccgcca tgcat
//
