LOCUS       sequence-354844-        5033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express iRFP with plasma membrane-targeting sequence in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5033)
  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 5033)
  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-iRFP-KRasCT
FEATURES             Location/Qualifiers
     source          1..5033
                     /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    1821..1942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1858..1877)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1912..1931
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1949..2404)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2086..2107)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2298..2317
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2431..2535
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2537..2894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2537..2728
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2560..2580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2745..2880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2929..3723
                     /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             2929..3723
                     /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(2983..3002)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3593..3612
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3911..3930)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3955..4002
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4331..4919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4820..4839
                     /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 agccatggaa ttcaccatgg ccgaagggag cgtggcaaga cagcccgatc
      661 tgctgacatg cgacgacgaa ccaatccaca ttccaggagc tatccagcct catggactgc
      721 tgctggctct ggccgctgac atgactatcg tggcaggctc tgataacctg ccagagctga
      781 ccgggctggc tatcggagca ctgattggcc ggtccgcagc cgacgtgttc gattctgaga
      841 ctcacaatag actgaccatc gccctggctg aacctggggc tgcagtggga gccccaatta
      901 ccgtcggctt cacaatgcga aaggacgctg gctttatcgg gtcctggcac cgccatgatc
      961 agctgatttt tctggagctg gaaccacctc agagagacgt ggcagagcca caggctttct
     1021 ttcggagaac aaacagcgcc atcaggcgcc tgcaggccgc tgagactctg gaatccgctt
     1081 gcgcagccgc tgcacaggaa gtgaggaaga tcaccgggtt cgatcgcgtc atgatctaca
     1141 gattcgcctc agactttagc ggagaagtga tcgcagaaga tcgctgtgcc gaggtcgaaa
     1201 gtaaactggg cctgcattac ccagcctcaa ccgtgcctgc acaggcacga cggctgtata
     1261 caatcaaccc cgtgcggatc attcccgaca ttaattacag accagtgccc gtcacacctg
     1321 acctgaaccc agtgactggg aggcccatcg atctgagctt cgccattctg cgcagcgtgt
     1381 cccctgtcca cctggagttt atgaggaata tcggaatgca tggcaccatg tctatcagta
     1441 ttctgagagg cgaaaggctg tgggggctga tcgtgtgcca ccatcggaca ccttactatg
     1501 tggacctgga cggcaggcag gcttgtgaac tggtggctca ggtgctggct tggcagattg
     1561 gcgtgatgga agaaagatct ggaggatccg gaggactcga gagcggccgc aagatgagca
     1621 aagatggtaa aaagaagaaa aagaagtcaa agacaaagtg tgtaattatg taagggccgc
     1681 tgactgactg aagatccacc ggatctagat aactgatcat aatcagccat accacatttg
     1741 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1801 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     1861 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1921 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     1981 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2041 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2101 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2161 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2221 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2281 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2341 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2401 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2461 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2521 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     2581 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2641 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2701 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2761 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2821 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2881 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2941 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3001 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3061 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     3121 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3181 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3241 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3301 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3361 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3421 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     3481 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3541 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3601 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3661 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3721 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3781 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3841 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     3901 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3961 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4021 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4081 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4141 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     4201 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4261 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4321 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4381 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4441 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4501 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4561 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4621 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4681 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4741 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4801 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4861 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4921 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4981 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
