LOCUS       sequence-354824-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express iRFP with golgi apparatus-targeting sequence in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  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 5296)
  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-GalT-iRFP
FEATURES             Location/Qualifiers
     source          1..5296
                     /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    2084..2205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2121..2140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2175..2194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2212..2667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2349..2370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2561..2580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2694..2798
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2800..3157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2800..2991
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2823..2843)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3008..3143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3192..3986
                     /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             3192..3986
                     /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(3246..3265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3856..3875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4174..4193)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4218..4265
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4594..5182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5083..5102
                     /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 gaggccacca tgaggcttcg ggagccgctc ctgagcggca
      661 gcgccgcgat gccaggcgcg tccctacagc gggcctgccg cctgctcgtg gccgtctgcg
      721 ctctgcacct tggcgtcacc ctcgtttact acctggctgg ccgcgacctg agccgcctgc
      781 cccaactggt cggagtctcc acaccgctgc agggcggctc gaacagtgcc gccgccatcg
      841 ggcagtcctc cggggagctc cggaccggag gggcctgcgg ccgcggagga tccggaggag
      901 aattcaccat ggccgaaggg agcgtggcaa gacagcccga tctgctgaca tgcgacgacg
      961 aaccaatcca cattccagga gctatccagc ctcatggact gctgctggct ctggccgctg
     1021 acatgactat cgtggcaggc tctgataacc tgccagagct gaccgggctg gctatcggag
     1081 cactgattgg ccggtccgca gccgacgtgt tcgattctga gactcacaat agactgacca
     1141 tcgccctggc tgaacctggg gctgcagtgg gagccccaat taccgtcggc ttcacaatgc
     1201 gaaaggacgc tggctttatc gggtcctggc accgccatga tcagctgatt tttctggagc
     1261 tggaaccacc tcagagagac gtggcagagc cacaggcttt ctttcggaga acaaacagcg
     1321 ccatcaggcg cctgcaggcc gctgagactc tggaatccgc ttgcgcagcc gctgcacagg
     1381 aagtgaggaa gatcaccggg ttcgatcgcg tcatgatcta cagattcgcc tcagacttta
     1441 gcggagaagt gatcgcagaa gatcgctgtg ccgaggtcga aagtaaactg ggcctgcatt
     1501 acccagcctc aaccgtgcct gcacaggcac gacggctgta tacaatcaac cccgtgcgga
     1561 tcattcccga cattaattac agaccagtgc ccgtcacacc tgacctgaac ccagtgactg
     1621 ggaggcccat cgatctgagc ttcgccattc tgcgcagcgt gtcccctgtc cacctggagt
     1681 ttatgaggaa tatcggaatg catggcacca tgtctatcag tattctgaga ggcgaaaggc
     1741 tgtgggggct gatcgtgtgc caccatcgga caccttacta tgtggacctg gacggcaggc
     1801 aggcttgtga actggtggct caggtgctgg cttggcagat tggcgtgatg gaagaaagat
     1861 ctctgactga ctgagatgcg cggatgtcga acggcagttc ggccgaatag gccgacagcc
     1921 ggccgtcggc cgacgacacg gccagggccg gcggggatcc accggatcta gataactgat
     1981 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2041 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2101 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2161 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     2221 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2281 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2341 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2401 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2461 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2521 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2581 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2641 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2701 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2761 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     2821 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2881 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3001 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3061 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3121 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3181 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3241 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3301 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3361 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3421 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3481 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3541 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3601 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3661 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3721 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3781 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3841 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3901 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3961 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4021 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4081 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4141 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4201 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4261 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4321 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4381 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4441 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4501 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4561 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4621 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4681 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4741 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4801 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4861 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4921 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4981 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5041 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5101 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5161 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5221 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5281 tattaccgcc atgcat
//
