LOCUS       sequence-254926-        4485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FAST (also called YFAST) fused to a mitochondria targeting
            sequence (mito) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4485)
  AUTHORS   Plamont MA, Billon-Denis E, Maurin S, Gauron C, Pimenta FM, Specht 
            CG, Shi J, Querard J, Pan B, Rossignol J, Moncoq K, Morellet N, 
            Volovitch M, Lescop E, Chen Y, Triller A, Vriz S, Le Saux T, Jullien
            L, Gautier A
  TITLE     Small fluorescence-activating and absorption-shifting tag for 
            tunable protein imaging in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):497-502. doi: 
            10.1073/pnas.1513094113. Epub 2015 Dec 28.
  PUBMED    26711992
REFERENCE   2  (bases 1 to 4485)
  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: 
            "10.1073/pnas.1513094113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-01-19- 19"; volume: "113"; issue: "3"; pages: "497-502"
COMMENT     Sequence Label: pAG156 Mito-FAST
FEATURES             Location/Qualifiers
     source          1..4485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             632..706
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      725..734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             731..1105
                     /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="MEHVAFGSEDIENTLAKMDDGQLDGLAFGAIQLDGDGNILQYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDSPEFYGKFKEGVASGNLNTMFEWMIPTSRGPTK
                     VKVHMKKALSGDSYWVFVKRV"
     CDS             1112..1141
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1278..1399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1315..1334)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1369..1388
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1406..1861)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1543..1564)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1755..1774
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1888..1992
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1994..2351
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1994..2185
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2017..2037)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2202..2337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2386..3180
                     /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             2386..3180
                     /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(2440..2459)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3050..3069
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3368..3387)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3412..3459
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3788..4376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4277..4296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      601 cgctaccgga ctcagatccg aattcgccac catgtccgtc ctgacgccgc tgctgctgcg
      661 gggcttgaca ggctcggccc ggcggctccc agtgccgcgc gccaagatcc attcgttgag
      721 atctgccacc atggagcatg ttgcctttgg cagtgaggac atcgagaaca ctctggccaa
      781 aatggacgac ggacaactgg atgggttggc ctttggcgca attcagctcg atggtgacgg
      841 gaatatcctg cagtacaatg ctgctgaagg agacatcaca ggcagagatc ccaaacaggt
      901 gattgggaag aacttcttca aggatgttgc acctggaacg gattctcccg agttttacgg
      961 caaattcaag gaaggcgtag cgtcagggaa tctgaacacc atgttcgaat ggatgatacc
     1021 gacaagcagg ggaccaacca aggtcaaggt gcacatgaag aaagcccttt ccggtgacag
     1081 ctattgggtc tttgtgaaac gggtgggatc cgaacaaaag cttatttctg aagaggactt
     1141 gtaatagccc gggtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     1201 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1261 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1321 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1381 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1441 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1501 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1561 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1621 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     1681 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     1741 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     1801 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     1861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1981 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2101 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2161 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2221 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2281 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2341 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2401 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2461 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2521 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     2581 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2641 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2701 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2761 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2821 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2881 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     2941 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3001 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3061 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3121 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3181 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3241 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3301 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3361 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3421 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3481 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3541 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3601 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     3661 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3901 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4441 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcca
//
