LOCUS       sequence-194701-        6452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of Pink1-YFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6452)
  AUTHORS   Narendra DP, Jin SM, Tanaka A, Suen DF, Gautier CA, Shen J, Cookson 
            MR, Youle RJ
  TITLE     PINK1 is selectively stabilized on impaired mitochondria to activate
            Parkin.
  JOURNAL   PLoS Biol. 2010 Jan 26;8(1):e1000298. doi: 
            10.1371/journal.pbio.1000298.
  PUBMED    20126261
REFERENCE   2  (bases 1 to 6452)
  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: "PLoS 
            Biol."; date: "2010-01-26"; volume: "8(1):e1000298. doi"; pages: " 
            10.1371/journal.pbio.1000298"
COMMENT     Sequence Label: pEYFP-N1-Pink1
FEATURES             Location/Qualifiers
     source          1..6452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1572..1581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1578..1580,1581..1583,1584..2297)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1623..1644)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1884..1903)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2231..2252
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2420..2541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2457..2476)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2511..2530
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2548..3003)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2685..2706)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2897..2916
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3030..3134
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3136..3493
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3136..3327
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3159..3179)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3344..3479
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3528..4322
                     /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             3528..4322
                     /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(3582..3601)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4192..4211
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4510..4529)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4554..4601
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4930..5518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5419..5438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5693..5996
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5997..6200
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6151..6171
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6260..6269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6266..6452,1..1556)
                     /codon_start=1
                     /product="human PTEN-induced putative kinase 1, a 
                     mitochondria serine/threonine protein kinase"
                     /label=PINK1
                     /translation="MAVRQALGRGLQLGRALLLRFTGKPGRAYGLGRPGPAAGCVRGER
                     PGWAAGPGAEPRRVGLGLPNRLRFFRQSVAGLAARLQRQFVVRAWGCAGPCGRAVFLAF
                     GLGLGLIEEKQAESRRAVSACQEIQAIFTQKSKPGPDPLDTRRLQGFRLEEYLIGQSIG
                     KGCSAAVYEATMPTLPQNLEVTKSTGLLPGRGPGTSAPGEGQERAPGAPAFPLAIKMMW
                     NISAGSSSEAILNTMSQELVPASRVALAGEYGAVTYRKSKRGPKQLAPHPNIIRVLRAF
                     TSSVPLLPGALVDYPDVLPSRLHPEGLGHGRTLFLVMKNYPCTLRQYLCVNTPSPRLAA
                     MMLLQLLEGVDHLVQQGIAHRDLKSDNILVELDPDGCPWLVIADFGCCLADESIGLQLP
                     FSSWYVDRGGNGCLMAPEVSTARPGPRAVIDYSKADAWAVGAIAYEIFGLVNPFYGQGK
                     AHLESRSYQEAQLPALPESVPPDVRQLVRALLQREASKRPSARVAANVLHLSLWGEHIL
                     ALKNLKLDKMVGWLLQQSAATLLANRLTEKCCVETKMKMLFLANLECETLCQAALLLCS
                     WRAAL"
ORIGIN
        1 tccctaaccg tctccgcttc ttccgccagt cggtggccgg gctggcggcg cggttgcagc
       61 ggcagttcgt ggtgcgggcc tggggctgcg cgggcccttg cggccgggca gtctttctgg
      121 ccttcgggct agggctgggc ctcatcgagg aaaaacaggc ggagagccgg cgggcggtct
      181 cggcctgtca ggagatccag gcaattttta cccagaaaag caagccgggg cctgacccgt
      241 tggacacgag acgcttgcag ggctttcggc tggaggagta tctgataggg cagtccattg
      301 gtaagggctg cagtgctgct gtgtatgaag ccaccatgcc tacattgccc cagaacctgg
      361 aggtgacaaa gagcaccggg ttgcttccag ggagaggccc aggtaccagt gcaccaggag
      421 aagggcagga gcgagctccg ggggcccctg ccttcccctt ggccatcaag atgatgtgga
      481 acatctcggc aggttcctcc agcgaagcca tcttgaacac aatgagccag gagctggtcc
      541 cagcgagccg agtggccttg gctggggagt atggagcagt cacttacaga aaatccaaga
      601 gaggtcccaa gcaactagcc cctcacccca acatcatccg ggttctccgc gccttcacct
      661 cttccgtgcc gctgctgcca ggggccctgg tcgactaccc tgatgtgctg ccctcacgcc
      721 tccaccctga aggcctgggc catggccgga cgctgttcct cgttatgaag aactatccct
      781 gtaccctgcg ccagtacctt tgtgtgaaca cacccagccc ccgcctcgcc gccatgatgc
      841 tgctgcagct gctggaaggc gtggaccatc tggttcaaca gggcatcgcg cacagagacc
      901 tgaaatccga caacatcctt gtggagctgg acccagacgg ctgcccctgg ctggtgatcg
      961 cagattttgg ctgctgcctg gctgatgaga gcatcggcct gcagttgccc ttcagcagct
     1021 ggtacgtgga tcggggcgga aacggctgtc tgatggcccc agaggtgtcc acggcccgtc
     1081 ctggccccag ggcagtgatt gactacagca aggctgatgc ctgggcagtg ggagccatcg
     1141 cctatgaaat cttcgggctt gtcaatccct tctacggcca gggcaaggcc caccttgaaa
     1201 gccgcagcta ccaagaggct cagctacctg cactgcccga gtcagtgcct ccagacgtga
     1261 gacagttggt gagggcactg ctccagcgag aggccagcaa gagaccatct gcccgagtag
     1321 ccgcaaatgt gcttcatcta agcctctggg gtgaacatat tctagccctg aagaatctga
     1381 agttagacaa gatggttggc tggctcctcc aacaatcggc cgccactttg ttggccaaca
     1441 ggctcacaga gaagtgttgt gtggaaacaa aaatgaagat gctctttctg gctaacctgg
     1501 agtgtgaaac gctctgccag gcagccctcc tcctctgctc atggagggca gccctggggg
     1561 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1621 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1681 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1741 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct
     1801 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1861 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1921 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1981 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     2041 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2101 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2161 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     2221 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2281 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     2341 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2401 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2461 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2521 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2581 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2641 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2701 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2761 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2821 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2881 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2941 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3001 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3061 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3121 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3241 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3301 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3361 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3421 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3481 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3541 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3601 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3661 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3721 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3781 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3841 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3901 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3961 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4021 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4081 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4141 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4201 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4261 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4321 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4381 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4441 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4501 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4561 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4621 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4681 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4741 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4801 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4861 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4921 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4981 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5041 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5101 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5161 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5221 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5281 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5341 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5401 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5461 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5521 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5581 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     5641 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     5701 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     5761 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5821 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     5881 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5941 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     6001 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     6061 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     6121 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     6181 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
     6241 agatctcgag ctcaagcttg ccaccatggc ggtgcgacag gcgctgggcc gcggcctgca
     6301 gctgggtcga gcgctgctgc tgcgcttcac gggcaagccc ggccgggcct acggcttggg
     6361 gcggccgggc ccggcggcgg gctgtgtccg cggggagcgt ccaggctggg ccgcaggacc
     6421 gggcgcggag cctcgcaggg tcgggctcgg gc
//
