LOCUS       sequence-192731-        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"
     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"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             634..2376
                     /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"
     regulatory      2392..2401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2398..2400,2401..2403,2404..3117)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2443..2464)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2704..2723)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3051..3072
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3240..3361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3277..3296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3331..3350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3368..3823)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3505..3526)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3717..3736
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3850..3954
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3956..4313
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3956..4147
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3979..3999)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4164..4299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4348..5142
                     /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             4348..5142
                     /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(4402..4421)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5012..5031
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5330..5349)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5374..5421
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5750..6338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6239..6258
                     /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 ccggactcag atctcgagct caagcttgcc accatggcgg tgcgacaggc gctgggccgc
      661 ggcctgcagc tgggtcgagc gctgctgctg cgcttcacgg gcaagcccgg ccgggcctac
      721 ggcttggggc ggccgggccc ggcggcgggc tgtgtccgcg gggagcgtcc aggctgggcc
      781 gcaggaccgg gcgcggagcc tcgcagggtc gggctcgggc tccctaaccg tctccgcttc
      841 ttccgccagt cggtggccgg gctggcggcg cggttgcagc ggcagttcgt ggtgcgggcc
      901 tggggctgcg cgggcccttg cggccgggca gtctttctgg ccttcgggct agggctgggc
      961 ctcatcgagg aaaaacaggc ggagagccgg cgggcggtct cggcctgtca ggagatccag
     1021 gcaattttta cccagaaaag caagccgggg cctgacccgt tggacacgag acgcttgcag
     1081 ggctttcggc tggaggagta tctgataggg cagtccattg gtaagggctg cagtgctgct
     1141 gtgtatgaag ccaccatgcc tacattgccc cagaacctgg aggtgacaaa gagcaccggg
     1201 ttgcttccag ggagaggccc aggtaccagt gcaccaggag aagggcagga gcgagctccg
     1261 ggggcccctg ccttcccctt ggccatcaag atgatgtgga acatctcggc aggttcctcc
     1321 agcgaagcca tcttgaacac aatgagccag gagctggtcc cagcgagccg agtggccttg
     1381 gctggggagt atggagcagt cacttacaga aaatccaaga gaggtcccaa gcaactagcc
     1441 cctcacccca acatcatccg ggttctccgc gccttcacct cttccgtgcc gctgctgcca
     1501 ggggccctgg tcgactaccc tgatgtgctg ccctcacgcc tccaccctga aggcctgggc
     1561 catggccgga cgctgttcct cgttatgaag aactatccct gtaccctgcg ccagtacctt
     1621 tgtgtgaaca cacccagccc ccgcctcgcc gccatgatgc tgctgcagct gctggaaggc
     1681 gtggaccatc tggttcaaca gggcatcgcg cacagagacc tgaaatccga caacatcctt
     1741 gtggagctgg acccagacgg ctgcccctgg ctggtgatcg cagattttgg ctgctgcctg
     1801 gctgatgaga gcatcggcct gcagttgccc ttcagcagct ggtacgtgga tcggggcgga
     1861 aacggctgtc tgatggcccc agaggtgtcc acggcccgtc ctggccccag ggcagtgatt
     1921 gactacagca aggctgatgc ctgggcagtg ggagccatcg cctatgaaat cttcgggctt
     1981 gtcaatccct tctacggcca gggcaaggcc caccttgaaa gccgcagcta ccaagaggct
     2041 cagctacctg cactgcccga gtcagtgcct ccagacgtga gacagttggt gagggcactg
     2101 ctccagcgag aggccagcaa gagaccatct gcccgagtag ccgcaaatgt gcttcatcta
     2161 agcctctggg gtgaacatat tctagccctg aagaatctga agttagacaa gatggttggc
     2221 tggctcctcc aacaatcggc cgccactttg ttggccaaca ggctcacaga gaagtgttgt
     2281 gtggaaacaa aaatgaagat gctctttctg gctaacctgg agtgtgaaac gctctgccag
     2341 gcagccctcc tcctctgctc atggagggca gccctggggg atccaccggt cgccaccatg
     2401 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2461 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2521 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2581 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     2641 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2701 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2761 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2821 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2881 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2941 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3001 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3061 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     3121 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3181 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3241 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3301 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3361 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3421 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3481 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3541 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3601 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3661 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3721 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3781 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     3841 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3901 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     3961 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4141 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4201 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4261 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4321 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4381 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4441 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4501 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     4561 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4621 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4681 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4741 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4801 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4861 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4921 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4981 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5041 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5101 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5161 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5221 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5281 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     5341 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     5401 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     5461 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     5521 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     5581 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5641 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5701 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5761 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5821 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5881 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5941 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6001 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6061 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6121 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6181 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6241 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6301 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6361 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6421 attctgtgga taaccgtatt accgccatgc at
//
