LOCUS       sequence-406794-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PINK1-mCherry-FIREtag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  AUTHORS   Bottone S, Joliot O, Cakil ZV, El Hajji L, Rakotoarison LM, 
            Boncompain G, Perez F, Gautier A
  TITLE     A fluorogenic chemically induced dimerization technology for 
            controlling, imaging and sensing protein proximity.
  JOURNAL   Nat Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8.
  PUBMED    37640938
REFERENCE   2  (bases 1 to 6117)
  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 
            Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8."
COMMENT     Sequence Label: pAG1238 PINK1-mCherry-FIREtag
FEATURES             Location/Qualifiers
     source          1..6117
                     /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             600..1754
                     /codon_start=1
                     /product="kinase domain from human PTEN-induced putative 
                     kinase 1, a mitochondria serine/threonine protein kinase"
                     /label=PINK1 kinase domain
                     /translation="EEKQAESRRAVSACQEIQAIFTQKSKPGPDPLDTRRLQGFRLEEY
                     LIGQSIGKGCSAAVYEATMPTLPQNLEVTKSTGLLPGRGPGTSAPGEGQERAPGAPAFP
                     LAIKMMWNISAGSSSEAILNTMSQELVPASRVALAGEYGAVTYRKSKRGPKQLAPHPNI
                     IRVLRAFTSSVPLLPGALVDYPDVLPSRLHPEGLGHGRTLFLVMKNYPCTLRQYLCVNT
                     PSPRLAAMMLLQLLEGVDHLVQQGIAHRDLKSDNILVELDPDGCPWLVIADFGCCLADE
                     SIGLQLPFSSWYVDRGGNGCLMAPEVSTARPGPRAVIDYSKADAWAVGAIAYEIFGLVN
                     PFYGQGKAHLESRSYQEAQLPALPESVPPDVRQLVRALLQREASK"
     CDS             2019..2726
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2164..2182)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2432..2451
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2905..3026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2942..2961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2996..3015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3033..3488)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3170..3191)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3382..3401
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3515..3619
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3621..3978
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3621..3812
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3644..3664)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3829..3964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4013..4807
                     /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             4013..4807
                     /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(4067..4086)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4677..4696
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4995..5014)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5039..5086
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5415..6003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5904..5923
                     /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 gctatgatcg
      601 aggaaaaaca ggcggagagc cggcgggcgg tctcggcctg tcaggagatc caggcaattt
      661 ttacccagaa aagcaagccg gggcctgacc cgttggacac gagacgcttg cagggctttc
      721 ggctggagga gtatctgata gggcagtcca ttggtaaggg ctgcagtgct gctgtgtatg
      781 aagccaccat gcctacattg ccccagaacc tggaggtgac aaagagcacc gggttgcttc
      841 cagggagagg cccaggtacc agtgcaccag gagaagggca ggagcgagct ccgggggccc
      901 ctgccttccc cttggccatc aagatgatgt ggaacatctc ggcaggttcc tccagcgaag
      961 ccatcttgaa cacaatgagc caggagctgg tcccagcgag ccgagtggcc ttggctgggg
     1021 agtatggagc agtcacttac agaaaatcca agagaggtcc caagcaacta gcccctcacc
     1081 ccaacatcat ccgggttctc cgcgccttca cctcttccgt gccgctgctg ccaggggccc
     1141 tggtcgacta ccctgatgtg ctgccctcac gcctccaccc tgaaggcctg ggccatggcc
     1201 ggacgctgtt cctcgttatg aagaactatc cctgtaccct gcgccagtac ctttgtgtga
     1261 acacacccag cccccgcctc gccgccatga tgctgctgca gctgctggaa ggcgtggacc
     1321 atctggttca acagggcatc gcgcacagag acctgaaatc cgacaacatc cttgtggagc
     1381 tggacccaga cggctgcccc tggctggtga tcgcagattt tggctgctgc ctggctgatg
     1441 agagcatcgg cctgcagttg cccttcagca gctggtacgt ggatcggggc ggaaacggct
     1501 gtctgatggc cccagaggtg tccacggccc gtcctggccc cagggcagtg attgactaca
     1561 gcaaggctga tgcctgggca gtgggagcca tcgcctatga aatcttcggg cttgtcaatc
     1621 ccttctacgg ccagggcaag gcccaccttg aaagccgcag ctaccaagag gctcagctac
     1681 ctgcactgcc cgagtcagtg cctccagacg tgagacagtt ggtgagggca ctgctccagc
     1741 gagaggccag caagagacca tctgcccgag tagccgcaaa tgtgcttcat ctaagcctct
     1801 ggggtgaaca tattctagcc ctgaagaatc tgaagttaga caagatggtt ggctggctcc
     1861 tccaacaatc ggccgccact ttgttggcca acaggctcac agagaagtgt tgtgtggaaa
     1921 caaaaatgaa gatgctcttt ctggctaacc tggagtgtga aacgctctgc caggcagccc
     1981 tcctcctctg ctcatggagg gcagccctgt ccggagggat ggtgagcaag ggcgaggagg
     2041 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     2101 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2161 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     2221 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     2281 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2341 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     2401 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     2461 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     2521 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2581 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2641 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2701 ccggcggcat ggacgagctg tacaagtcta gaggcggcgg ctccggtgac agatattggg
     2761 tctttgtgaa acgggtgtaa tgagcggccg cgactctaga tcataatcag ccataccaca
     2821 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2881 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     2941 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3001 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3061 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3121 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3181 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3241 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3301 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3361 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3421 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3481 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3541 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3601 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3661 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3721 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3781 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3841 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3901 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3961 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4021 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4081 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4141 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4201 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4261 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4321 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4381 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4441 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4501 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4561 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4621 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4681 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4741 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4801 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4861 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4921 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4981 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5041 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5101 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5161 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5221 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5281 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5341 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5521 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5761 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5821 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5881 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5941 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6001 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6061 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
