LOCUS       sequence-354718-        5586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express iRFP with late endosome-targeting sequence in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  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 5586)
  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-Rab7
FEATURES             Location/Qualifiers
     source          1..5586
                     /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    2374..2495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2411..2430)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2465..2484
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2502..2957)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2639..2660)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2851..2870
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2984..3088
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3090..3447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3090..3281
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3113..3133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3298..3433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3482..4276
                     /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             3482..4276
                     /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(3536..3555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4146..4165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4464..4483)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4508..4555
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4884..5472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5373..5392
                     /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 aagtactgaa 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 gatgacctct aggaagaaag
     1621 tgttgctgaa ggtcatcatc ctgggggact ctggtgttgg aaagacctct ctcatgaacc
     1681 agtatgtgaa caagaagttc agtaaccagt acaaagccac aataggagcg gactttctga
     1741 ccaaggaggt gatggtggac gacagacttg ttaccatgca gatctgggac acagccggtc
     1801 aagaacggtt ccagtctctt ggtgtggcct tctacagagg tgcagattgc tgtgttctgg
     1861 tgtttgatgt gactgccccc aacactttca aaaccctcga cagctggaga gacgagtttc
     1921 tcatccaggc cagcccccgg gatcccgaga acttcccttt tgttgtgttg ggaaacaaga
     1981 ttgacctgga aaacagacaa gtggccacaa agagggcaca ggcttggtgc tacagcaaaa
     2041 acaacattcc ttacttcgag accagtgcca aggaggccat caatgtggag caggccttcc
     2101 agacaattgc tcggaatgcc cttaaacagg aaacagaagt ggaactgtac aatgaattcc
     2161 ctgaacccat caaactggac aagaatgacc gggccaaggc ctccgcagaa agctgcagtt
     2221 gttgagcggc cgctgactga ctgaagatcc accggatcta gataactgat cataatcagc
     2281 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2341 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2401 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2461 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2521 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2581 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2641 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2701 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2761 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2821 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2881 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2941 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3001 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3061 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3121 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3181 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3241 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3301 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3361 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3421 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3481 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3541 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3601 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3661 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3721 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3781 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3841 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3901 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3961 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4021 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4081 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4141 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4201 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4261 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4321 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4381 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4441 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4501 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4561 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4621 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4681 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4741 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4801 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4861 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4921 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4981 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5041 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5101 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5161 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5221 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5281 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5341 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5401 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5461 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5521 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5581 atgcat
//
