LOCUS       sequence-354793-        5619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express iRFP with recycling endosome-targeting sequence in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5619)
  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 5619)
  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-Rab11
FEATURES             Location/Qualifiers
     source          1..5619
                     /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"
     regulatory      1602..1611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2407..2528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2444..2463)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2498..2517
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2535..2990)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2672..2693)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2884..2903
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3017..3121
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3123..3480
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3123..3314
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3146..3166)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3331..3466
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3515..4309
                     /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             3515..4309
                     /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(3569..3588)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4179..4198
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4497..4516)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4541..4588
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4917..5505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5406..5425
                     /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 ggccaccatg ggcacccgcg
     1621 acgacgagta cgactacctc tttaaagttg tccttattgg agattctggt gttgggaaga
     1681 gtaacctcct gtctcgattt actcggaatg agtttaatct ggaaagcaag agtaccattg
     1741 gagtagaatt tgcaacaaga agcatccagg ttgatgggaa aacaataaag gcacagatat
     1801 gggacacagc agggcaggag cggtacaggg ctataacgtc tgcatactat cgtggagcag
     1861 taggtgcctt attggtttat gacattgcta agcatctcac atatgaaaat gtagaacgat
     1921 ggctgaaaga actgagagat catgctgata gtaacattgt tatcatgctt gtgggcaata
     1981 agagtgattt acgtcatctc agggcagttc ctacagatga agcaagagca tttgcagaga
     2041 agaatggttt gtcgttcatt gagacatctg ctctagattc tacaaatgtt gaagctgctt
     2101 ttcagacaat tctaacagag atataccgca ttgtttctca gaagcaaatg tcagacagac
     2161 gtgaaaatga catgtctcca agcaacaatg cggttcctat tcatgtccca cccaccactg
     2221 agaacaagcc aaaggtgcag tgctgtcaga acatctaagc ggccgctgac tgactgaaga
     2281 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2341 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2401 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2461 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2521 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2581 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2641 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2701 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2761 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2821 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2881 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2941 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3001 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3061 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3121 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3301 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3361 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3421 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3481 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3541 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3601 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3661 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3721 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3781 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3841 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3901 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3961 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4021 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4081 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4141 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4201 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4261 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4321 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4381 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4441 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4501 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4561 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4621 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4681 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4741 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4801 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4861 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4921 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4981 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5041 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5101 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5161 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5221 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5281 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5341 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5401 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5461 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5521 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5581 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
