LOCUS       sequence-353705-        4476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mitochondrial targeting domain-pFAST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4476)
  AUTHORS   Benaissa H, Ounoughi K, Aujard I, Fischer E, Goiame R, Nguyen J, 
            Tebo AG, Li C, Le Saux T, Bertolin G, Tramier M, Danglot L, 
            Pietrancosta N, Morin X, Jullien L, Gautier A
  TITLE     Engineering of a fluorescent chemogenetic reporter with tunable 
            color for advanced live-cell imaging.
  JOURNAL   Nat Commun. 2021 Nov 30;12(1):6989. doi: 10.1038/s41467-021-27334-0.
  PUBMED    34848727
REFERENCE   2  (bases 1 to 4476)
  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"; journalName: "Nat 
            Commun."; date: "2021-11-30"; volume: "12(1):6989. doi"; pages: " 
            10.1038/s41467-021-27334-0"
COMMENT     Sequence Label: pAG671 mito-pFAST
FEATURES             Location/Qualifiers
     source          1..4476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        106..409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        410..613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     564..584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             672..746
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      765..774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             771..1145
                     /codon_start=1
                     /product="small protein tag, derived from photoactive 
                     yellow protein, that allows in vivo fluorescent labeling by
                     addition of a cell-permeant fluorogen (Plamont et al., 
                     2016)"
                     /label=Y-FAST
                     /translation="MEHVAFGSEDIENTLANMDDEQLDRLAFGVIQLDGDGNILLYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDTPEFYGKFKEGAASGNLNTMFEWTIPTSRGPTK
                     VKVHLKKALSGDRYWVFVKRV"
     CDS             1152..1181
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1309..1430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1346..1365)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1400..1419
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1437..1892)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1574..1595)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1786..1805
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1919..2023
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2025..2382
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2025..2216
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2048..2068)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2233..2368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2417..3211
                     /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             2417..3211
                     /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(2471..2490)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3081..3100
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3399..3418)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3443..3490
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3819..4407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4308..4327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
       61 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      421 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      481 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      541 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      601 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatccg
      661 aattcgccac catgtccgtc ctgacgccgc tgctgctgcg gggcttgaca ggctcggccc
      721 ggcggctccc agtgccgcgc gccaagatcc attcgttgag atctgccacc atggagcatg
      781 ttgcctttgg cagtgaggac atcgagaaca ctctggccaa tatggacgac gaacaactgg
      841 ataggttggc ctttggcgta attcagctcg atggtgacgg gaatatcctg ctgtacaatg
      901 ctgctgaagg ggacatcact ggcagagatc ccaaacaggt gattgggaag aacttcttca
      961 aggatgttgc acctggaacg gatactcccg agttttacgg caaattcaag gaaggcgcag
     1021 cgtcagggaa tctgaacacc atgttcgaat ggacgatacc gacaagcagg ggaccaacca
     1081 aggtcaaggt gcacttgaag aaagcccttt ccggtgacag atattgggtc tttgtgaaac
     1141 gggtgggatc cgaacaaaag cttatttctg aagaggactt gtaataggcg gccgcgactc
     1201 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1261 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1321 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1381 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     1441 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1501 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1561 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1621 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1681 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1741 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1801 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     1861 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     1921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1981 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2041 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2101 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2161 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2221 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2281 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2341 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     2401 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2461 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2521 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2581 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2641 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2701 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2761 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2821 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     2881 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2941 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3001 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3061 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3121 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3181 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3241 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3301 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3361 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     3421 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     3481 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3541 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3601 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     3661 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3721 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3781 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3961 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4381 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4441 gccttttgct ggccttttgc tcacatgttc tttcct
//
