LOCUS       sequence-354691-        5610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express iRFP with early endosome-targeting sequence in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5610)
  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 5610)
  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-Rab5
FEATURES             Location/Qualifiers
     source          1..5610
                     /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    2398..2519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2435..2454)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2489..2508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2526..2981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2663..2684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2875..2894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3008..3112
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3114..3471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3114..3305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3137..3157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3322..3457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3506..4300
                     /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             3506..4300
                     /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(3560..3579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4170..4189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4488..4507)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4532..4579
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4908..5496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5397..5416
                     /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 gatggctaat cgaggagcaa
     1621 caagacccaa cgggccaaat actggaaata aaatatgcca gttcaaactg gtccttctag
     1681 gagagtctgc tgttggcaaa tcaagcctgg ttcttcgctt tgtgaaaggc caatttcatg
     1741 aatttcaaga gagtaccatt ggggctgcct ttctaaccca aactgtgtgt cttgatgaca
     1801 caacagtaaa atttgaaata tgggatacag ctggtcaaga acggtatcat agcttagcac
     1861 caatgtacta ccgaggagca caagcagcca tagttgtgta tgatatcaca aatgaggaat
     1921 cctttgcgag agcaaaaaac tgggttaaag aacttcaaag gcaagcaagt cctaatattg
     1981 tgatagcttt gtcaggaaac aaagctgact tagcaaataa aagagctgtt gacttccagg
     2041 aagcacagtc ctatgcagat gacaacagct tattatttat ggagacatca gctaagacat
     2101 caatgaatgt aaatgaaata tttatggcaa tagctaaaaa gctgccaaag aatgaaccac
     2161 agaatcctgg tgcaaactca gccagaggac gaggagtaga ccttactgag cctgcacagc
     2221 cagccagaag ccagtgttgt agtaactgag cggccgctga ctgactgaag atccaccgga
     2281 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2341 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2401 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2461 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2521 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2581 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2641 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2701 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2761 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2821 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2881 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2941 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3001 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3121 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3481 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3541 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3601 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3661 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3721 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3781 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3841 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3901 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3961 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4021 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4081 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4141 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4201 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4261 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4321 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4381 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4441 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4501 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4561 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4621 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4681 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4741 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4801 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4861 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4921 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4981 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5041 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5101 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5161 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5221 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5281 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5341 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5401 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5461 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5521 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5581 tctgtggata accgtattac cgccatgcat
//
