LOCUS       sequence-469931-        9181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus plasmid for the stable expression of mCherry-Parkin in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9181)
  AUTHORS   Rose K, Herrmann E, Kakudji E, Lizarrondo J, Celebi AY, Wilfling F, 
            Lewis SC, Hurley JH
  TITLE     In situ cryo-ET visualization of mitochondrial depolarization and 
            mitophagic engulfment.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Aug 5;122(31):e2511890122. doi: 
            10.1073/pnas.2511890122. Epub 2025 Jul 31.
  PUBMED    40743392
REFERENCE   2  (bases 1 to 9181)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-08-5"; volume: "122(31):e2511890122. 
            doi"; pages: " 10.1073/pnas.2511890122. Epub 2025 Jul 31"
COMMENT     Sequence Label: pLV-mCherry-Parkin
FEATURES             Location/Qualifiers
     source          1..9181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        209..313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(314..382,383..1174)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(532..551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1345..1933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1834..1853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             1995..2175
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     2684..2700
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2685..2701
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2704..3292
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2810..2821
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3219..3239
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4819..5526)
                     /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(5094..5113)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5363..5381
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5523..5532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          5548..6486
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5571..5591)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        complement(6487..6698)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    6801..6918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7216..7260)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7445..7678
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8175..8300
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             8347..8527
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8545..8743)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8570..8590)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        8745..9124
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc
       61 actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat
      121 cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac
      181 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      241 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      301 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      361 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      421 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      481 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      541 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
      601 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      661 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
      721 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
      781 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
      841 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
      901 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
      961 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1021 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1081 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1141 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1201 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1261 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1321 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1381 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1441 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1501 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1561 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1621 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1681 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1741 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1801 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1861 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1921 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1981 ttttgctcac atgctgctag agattttcca cactgactaa aagggtctga gggatctcta
     2041 gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta
     2101 ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc
     2161 taaccagaga gacccagtac agtccggatg cagctctcgg gccatgtgat gaaatgctag
     2221 gcggctgtca aacctccact ctaatacttc tctctccggg tcatccatcc catgcaggct
     2281 cacagggtgt aacaagcggg tgttctctcc ttcattggct tcttctacct tctcttgctc
     2341 aactggtact agcttgtagc accatccaaa ggtcagtgga tatctgatcc ctggccctgg
     2401 tgtgtagttc tgccaatcag ggaagtagcc ttgtgtgtgg tagatccaca gatcaaggat
     2461 atcttgtctt cgttgggagt gaattagccc ttccagtccc cccttttctt ttaaaaagtg
     2521 gctaagatct acagctgcct tgtaagtcat tggtcttaaa ggtacctgag gtgtgactgg
     2581 aaaacccacc tcctcctcct cttgtgcttc tagccaggca caatcagcat tggtagctgc
     2641 tgtattgcta cttgtgattg ctccatgttt ttctaggtct cgatcgaggt cgacggtatc
     2701 gatgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg
     2761 gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg
     2821 aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc
     2881 aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag
     2941 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg
     3001 ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg
     3061 ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct
     3121 cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     3181 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     3241 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa
     3301 gcttgatatc gaattgtacc tagtggaacc ggaaccctta aacatgtctc gacctggctt
     3361 actagtcggt tcaactctag gtttacttgt acactacacg tcgaaccagt ggtcccccat
     3421 gcagacgcgg ttccactcgc agccacagtt ccagcaccac tcgagcctgc actggggctg
     3481 cggacacttc atgtgcatgc agcctccatt tttttccact ggtacatggc agcggggaca
     3541 gggcttggtg gttttcttga tggtttcttt ggaggctgct tcccaacgag cctgctcggc
     3601 ggctctttca tcgactctgt aggcctgagt agttgttcct gaggcttcaa atacggcact
     3661 gcactcccct tcatggtacg cttctttaca ttcccggcag aaggcaaacc cacagcccag
     3721 gccattgccc ccttcgcagg tgactttcct ctggtcaggc tccggcagca gccccgctcc
     3781 acagccaggg cgggggcata acacgccccc catctgcagg acacactcct ctgcaccata
     3841 ctgctggtac cggttgtact gctcttctcc cagaatcctg aagtgatgga gctctttaat
     3901 caaggagttg ggacagccag ccacacaagg cagggagtag ccaagctgag ggtcgtggac
     3961 aaactgccga tcattgagtc ttgtcacaca gtataagtgg aaacagtcta agcaaatcac
     4021 gtggcgggag ttgcactgga aaaccaggac ggggctcctg acgtctgtgc acgtaatgca
     4081 agtgatgttc cgactatttg ttgcgatcag gtgcaaagct actgatgttt ccttgtcaga
     4141 ggtggggtgt gctccacatt taaagaaaaa ttctgcacta gtcccagggc agtgtgggga
     4201 ttggcattca ccactcatcc ggtttggaat taaaacatca tcccagcaag atggaccctg
     4261 ggtcaaggtg agcgttgcct gcctgcaggt gctgcactgt accctgagtt ttcccggctg
     4321 cactctttga caggggcctt tgcaatacac ataaaagctg ttgtagattg atctacctgc
     4381 tggacttcca gctggtggtg agtccttcct gctgtcagtg tgcagaatga cagccagccc
     4441 cacagagtct cctgggagga ctgagctgct gaggtccacc cgagtcaagc tctggggctc
     4501 ccgctcacag cctcccgccg cgtttctggg gtcgtcgcct ccagttgcat tcatttcttg
     4561 accttttctc cacggtctct gcacaatgtg aacaatgctc tgctgatcca ggtcacaatt
     4621 ctgcacagtc cagtcattcc tcagctcctt ccctgcgaaa atcacacgca actggtcagc
     4681 cggaaccccc tgtcgcttag caaccacctc cttgagctgg aagatgctgg tgtcagaatc
     4741 gacctccact gggaaaccat ggctggagtt gaacctgaca aacactatca aagcttgagc
     4801 tcgagatctg agtccggact tgtacagctc gtccatgccg ccggtggagt ggcggccctc
     4861 ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt
     4921 gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt
     4981 gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt
     5041 cagggcgccg tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt
     5101 ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa
     5161 ctcgccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
     5221 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc
     5281 ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca
     5341 ggcgaagggc agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta
     5401 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat
     5461 gtgcaccttg aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct
     5521 caccatggtg gcgctagctc acgacacctg aaatggaaga aaaaaacttt gaaccactgt
     5581 ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca aggagaatta
     5641 catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg gaaactccat
     5701 cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg agaactaatc
     5761 gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg ctgaggacgg
     5821 aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc gccgcgtcct
     5881 ccattttgag ctccctgcag cagggccggg aagcggccat ctttccgctc acgcaactgg
     5941 tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg cgaggccagg
     6001 caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg gccgcgctcg
     6061 caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc cgcccgcggc
     6121 cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac tatcttgcca
     6181 gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact tatcgaaagc
     6241 agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc gcccaggcca
     6301 ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg aactctccca
     6361 cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg tggaagtaat
     6421 tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac cacacacggc
     6481 acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc
     6541 acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact
     6601 ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga cccctccccc
     6661 caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac atgcatggcg
     6721 gtaatacggt tatccacgcg gccctagagc gggcccggta ctaaccaaac tggatctctg
     6781 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
     6841 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
     6901 cccccttttc ttttaaaatt gtggatgaat actgccattt gtctgcagaa ttggcgcacg
     6961 cagtgccgat ccgttcacta atcgaatgga tctgtctctg tctctctctc caccttcttc
     7021 ttctattcct tcgggcctgt cgggtcccct cggggttggg aggtgggtct gaaacgataa
     7081 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     7141 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     7201 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     7261 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     7321 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     7381 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     7441 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     7501 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     7561 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     7621 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     7681 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     7741 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     7801 atctcctcct ccaggtctga agatcagcgg ccggccgctt gctgtgcggt ggtcttactt
     7861 ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt
     7921 tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct
     7981 gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac
     8041 aggccaggat taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat
     8101 ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc
     8161 ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa
     8221 atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc
     8281 cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg
     8341 cgccactgct agagattttc cacactgact aaaagggtct gagggatctc tagttaccag
     8401 agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct
     8461 taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga
     8521 gagacccagt acaggcaaaa cgcgctgctt atatagacct cccaccgtac acgcctaccg
     8581 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     8641 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     8701 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg
     8761 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     8821 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     8881 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     8941 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     9001 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     9061 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     9121 tgtcaacgcg tatatctggc ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc
     9181 a
//
