LOCUS       sequence-354834-        5272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SECFP with mitochondria-targeting sequence in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5272)
  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 5272)
  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-Tom20-SECFP
FEATURES             Location/Qualifiers
     source          1..5272
                     /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"
     CDS             join(1101..1103,1104..1106,1107..1817)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIGFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1146..1167)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1407..1426)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1754..1775
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2060..2181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2097..2116)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2151..2170
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2188..2643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2325..2346)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2537..2556
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2670..2774
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2776..3133
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2776..2967
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2799..2819)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2984..3119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3168..3962
                     /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             3168..3962
                     /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(3222..3241)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3832..3851
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4150..4169)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4194..4241
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4570..5158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5059..5078
                     /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 agccatgctc gagcccgcca tggtgggtcg gaacagcgcc atcgccgccg
      661 gtgtatgcgg ggcccttttc attgggtact gcatctactt cgaccgcaaa agacgaagtg
      721 accccaactt caagaacagg cttcgagaac gaagaaagaa acagaagctt gccaaggaga
      781 gagctgggct ttccaagtta cctgacctta aagatgctga agctgttcag aagttcttcc
      841 ttgaagaaat acagcttggt gaagagttac tagctcaagg tgaatatgag aagggcgtag
      901 accatctgac aaatgcaatt gctgtgtgtg gacagccaca gcagttactg caggtcttac
      961 agcaaactct tccaccacca gtgttccaga tgcttctgac taagctccca acaattagtc
     1021 agagaattgt aagtgctcag agcttggctg aagatgatgt ggaaggcggc cgcggaggat
     1081 ccggaggaga attcggatcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1141 ccatcctggt cgagctggac ggcgacgtaa acggccacag gttcagcgtg tccggcgagg
     1201 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1261 tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc
     1321 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1381 tccaggagcg taccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1441 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcggc ttcaaggagg
     1501 acggcaacat cctggggcac aagctagagt acaactacat cagccacaac gtctatatca
     1561 ccgccgacaa gcagaagaac ggcatcaagg cccacttcaa gatccgccac aacatcgagg
     1621 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1681 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1741 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1801 tggacgagct gtacaaggga ggatctggag gaagatctct gactgactga gatgcgcgga
     1861 tgtcgaacgg cagttcggcc gaataggccg acagccggcc gtcggccgac gacacggcca
     1921 gggccggcgg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     1981 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2041 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2101 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2161 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2221 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2281 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2341 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2401 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2461 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2521 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2581 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2641 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2701 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2761 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     2821 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2881 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2941 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3001 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3061 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3121 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3181 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3241 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3301 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3361 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3421 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3481 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3541 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3601 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3661 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     3721 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3781 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3841 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3901 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3961 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4021 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4081 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4141 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4201 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4261 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4321 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4381 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4441 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4501 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4561 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4621 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4681 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4741 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4801 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4861 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4921 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4981 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5041 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5101 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5161 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5221 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
