LOCUS       sequence-354826-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express iRFP with nucleus-targeting sequence in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  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 5422)
  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-H2B-iRFP
FEATURES             Location/Qualifiers
     source          1..5422
                     /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             624..1001
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     polyA_signal    2210..2331
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2247..2266)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2301..2320
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2338..2793)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2475..2496)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2687..2706
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2820..2924
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2926..3283
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2926..3117
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2949..2969)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3134..3269
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3318..4112
                     /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             3318..4112
                     /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(3372..3391)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3982..4001
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4300..4319)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4344..4391
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4720..5308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5209..5228
                     /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 aagtactctc gagatgccag agccagcgaa gtctgctccc gccccgaaaa
      661 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
      721 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
      781 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc
      841 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca
      901 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
      961 ccgagggtac taaggccatc accaagtaca ccagcgctaa gagcggccgc ggaggatccg
     1021 gaggagaatt caccatggcc gaagggagcg tggcaagaca gcccgatctg ctgacatgcg
     1081 acgacgaacc aatccacatt ccaggagcta tccagcctca tggactgctg ctggctctgg
     1141 ccgctgacat gactatcgtg gcaggctctg ataacctgcc agagctgacc gggctggcta
     1201 tcggagcact gattggccgg tccgcagccg acgtgttcga ttctgagact cacaatagac
     1261 tgaccatcgc cctggctgaa cctggggctg cagtgggagc cccaattacc gtcggcttca
     1321 caatgcgaaa ggacgctggc tttatcgggt cctggcaccg ccatgatcag ctgatttttc
     1381 tggagctgga accacctcag agagacgtgg cagagccaca ggctttcttt cggagaacaa
     1441 acagcgccat caggcgcctg caggccgctg agactctgga atccgcttgc gcagccgctg
     1501 cacaggaagt gaggaagatc accgggttcg atcgcgtcat gatctacaga ttcgcctcag
     1561 actttagcgg agaagtgatc gcagaagatc gctgtgccga ggtcgaaagt aaactgggcc
     1621 tgcattaccc agcctcaacc gtgcctgcac aggcacgacg gctgtataca atcaaccccg
     1681 tgcggatcat tcccgacatt aattacagac cagtgcccgt cacacctgac ctgaacccag
     1741 tgactgggag gcccatcgat ctgagcttcg ccattctgcg cagcgtgtcc cctgtccacc
     1801 tggagtttat gaggaatatc ggaatgcatg gcaccatgtc tatcagtatt ctgagaggcg
     1861 aaaggctgtg ggggctgatc gtgtgccacc atcggacacc ttactatgtg gacctggacg
     1921 gcaggcaggc ttgtgaactg gtggctcagg tgctggcttg gcagattggc gtgatggaag
     1981 aaagatctct gactgactga gatgcgcgga tgtcgaacgg cagttcggcc gaataggccg
     2041 acagccggcc gtcggccgac gacacggcca gggccggcgg ggatccaccg gatctagata
     2101 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2161 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2221 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2281 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     2341 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2401 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2461 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2521 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2581 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2641 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2701 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2761 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     2821 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2881 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     2941 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3001 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3061 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3121 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3181 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3241 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     3301 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3361 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3421 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3481 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3541 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3601 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3661 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3721 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3781 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3841 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3901 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3961 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4021 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4081 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4141 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4201 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4261 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     4321 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     4381 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4441 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4501 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     4561 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4861 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5101 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5401 taaccgtatt accgccatgc at
//
