LOCUS       sequence-75569-e        6087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human Park2 fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6087)
  AUTHORS   Narendra D, Tanaka A, Suen DF, Youle RJ
  TITLE     Parkin is recruited selectively to impaired mitochondria and 
            promotes their autophagy.
  JOURNAL   J Cell Biol. 2008 Dec 1. 183(5):795-803.
  PUBMED    19029340
REFERENCE   2  (bases 1 to 6087)
  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: "J Cell 
            Biol. 2008 Dec 1. 183(5):795-803."
COMMENT     Sequence Label: mCherry-Parkin
FEATURES             Location/Qualifiers
     source          1..6087
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2875..2996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2912..2931)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2966..2985
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3003..3458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3140..3161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3352..3371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3485..3589
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3591..3948
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3591..3782
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3614..3634)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3799..3934
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3983..4777
                     /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             3983..4777
                     /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(4037..4056)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4647..4666
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4965..4984)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5009..5056
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5385..5973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5874..5893
                     /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 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc tcaagctttg atagtgtttg tcaggttcaa ctccagccat
     1381 ggtttcccag tggaggtcga ttctgacacc agcatcttcc agctcaagga ggtggttgct
     1441 aagcgacagg gggttccggc tgaccagttg cgtgtgattt tcgcagggaa ggagctgagg
     1501 aatgactgga ctgtgcagaa ttgtgacctg gatcagcaga gcattgttca cattgtgcag
     1561 agaccgtgga gaaaaggtca agaaatgaat gcaactggag gcgacgaccc cagaaacgcg
     1621 gcgggaggct gtgagcggga gccccagagc ttgactcggg tggacctcag cagctcagtc
     1681 ctcccaggag actctgtggg gctggctgtc attctgcaca ctgacagcag gaaggactca
     1741 ccaccagctg gaagtccagc aggtagatca atctacaaca gcttttatgt gtattgcaaa
     1801 ggcccctgtc aaagagtgca gccgggaaaa ctcagggtac agtgcagcac ctgcaggcag
     1861 gcaacgctca ccttgaccca gggtccatct tgctgggatg atgttttaat tccaaaccgg
     1921 atgagtggtg aatgccaatc cccacactgc cctgggacta gtgcagaatt tttctttaaa
     1981 tgtggagcac accccacctc tgacaaggaa acatcagtag ctttgcacct gatcgcaaca
     2041 aatagtcgga acatcacttg cattacgtgc acagacgtca ggagccccgt cctggttttc
     2101 cagtgcaact cccgccacgt gatttgctta gactgtttcc acttatactg tgtgacaaga
     2161 ctcaatgatc ggcagtttgt ccacgaccct cagcttggct actccctgcc ttgtgtggct
     2221 ggctgtccca actccttgat taaagagctc catcacttca ggattctggg agaagagcag
     2281 tacaaccggt accagcagta tggtgcagag gagtgtgtcc tgcagatggg gggcgtgtta
     2341 tgcccccgcc ctggctgtgg agcggggctg ctgccggagc ctgaccagag gaaagtcacc
     2401 tgcgaagggg gcaatggcct gggctgtggg tttgccttct gccgggaatg taaagaagcg
     2461 taccatgaag gggagtgcag tgccgtattt gaagcctcag gaacaactac tcaggcctac
     2521 agagtcgatg aaagagccgc cgagcaggct cgttgggaag cagcctccaa agaaaccatc
     2581 aagaaaacca ccaagccctg tccccgctgc catgtaccag tggaaaaaaa tggaggctgc
     2641 atgcacatga agtgtccgca gccccagtgc aggctcgagt ggtgctggaa ctgtggctgc
     2701 gagtggaacc gcgtctgcat gggggaccac tggttcgacg tgtaaggatc caccggatct
     2761 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2821 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2881 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2941 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3001 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3061 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3481 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3541 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3601 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3661 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3721 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3781 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3841 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3901 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3961 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4021 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4081 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4141 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4201 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4261 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4321 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4381 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4441 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4501 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4561 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4621 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4681 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4741 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4801 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4861 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4921 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4981 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5041 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5101 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5161 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5221 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5281 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5341 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5401 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5461 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5521 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5581 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5641 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5701 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5761 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5821 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5881 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5941 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6001 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6061 gtggataacc gtattaccgc catgcat
//
