LOCUS       sequence-420145-        8467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for stable low expression of Parkin, internally 
            tagged with mKO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8467)
  AUTHORS   Waters CS, Angenent SB, Altschuler SJ, Wu LF
  TITLE     A PINK1 input threshold arises from positive feedback in the 
            PINK1/Parkin mitophagy decision circuit.
  JOURNAL   Cell Rep. 2023 Oct 31;42(10):113260. doi: 
            10.1016/j.celrep.2023.113260. Epub 2023 Oct 17.
  PUBMED    37851575
REFERENCE   2  (bases 1 to 8467)
  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: "Cell Rep.";
            date: "2023-10-31"; volume: "42(10):113260. doi"; pages: " 
            10.1016/j.celrep.2023.113260. Epub 2023 Oct 17"
COMMENT     Sequence Label: pLv-CMV(trunc)-Parkin-A92mKO2
FEATURES             Location/Qualifiers
     source          1..8467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..122
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     73..93
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             458..1108
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             458..1108
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     misc_feature    2274..2862
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2327..2347)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2745..2756)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3069..3702
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3830..3846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3830..3846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3843..3865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3854..3870
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3878..3884,3885..3902,3903..3908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3923..3944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4061..4078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4232..4820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4312..4331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4991..5782,5783..5851))
                     /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     5614..5633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5852..5956)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6004..6138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6041..6060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6095..6114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6306..6939
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6986..7111
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7608..7841
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8026..8070
                     /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    8333..8448
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
ORIGIN
        1 agcagagatc cagtttatcg atccaaaatc aacgggactt tccaaaatgt cgtaacaact
       61 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      121 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatga tagtgtttgt
      181 caggttcaac tccagccatg gtttcccagt ggaggtcgat tctgacacca gcatcttcca
      241 gctcaaggag gtggttgcta agcgacaggg ggttccggct gaccagttgc gtgtgatttt
      301 cgcagggaag gagctgagga atgactggac tgtgcagaat tgtgacctgg atcagcagag
      361 cattgttcac attgtgcaga gaccgtggag aaaaggtcaa gaaatgaatg caactggagg
      421 cgacgacccc agaaacgcgg cgggtgctgg ttctggcgtg agcgtgatca agcccgagat
      481 gaagatgagg tactacatgg acggctccgt caatgggcat gagttcacaa tcgagggtga
      541 gggcacaggc agaccttacg agggacatca ggagatgaca ctgcgcgtca caatggccga
      601 gggcgggcca atgcctttcg ccttcgacct ggtgtcccac gtgttctgtt acggccacag
      661 agtttttacc aagtacccag aagagatccc agactatttc aagcaggcct ttcctgaggg
      721 cctgtcctgg gagaggtccc tggagttcga ggacggcggc tccgcctccg tgagcgccca
      781 catcagcctg aggggcaaca ccttctacca caagtccaag ttcaccggcg tgaacttccc
      841 cgccgacggc cccatcatgc agaaccagag cgtggactgg gagccctcca ccgagaagat
      901 caccgccagc gacggcgtgc tgaagggcga cgtgaccatg tacctgaagc tggagggcgg
      961 cggcaaccac aagtgccaga tgaagaccac ctacaaggcc gccaaggaga tcctggagat
     1021 gcccggcgac cactacatcg gccacaggct ggtgaggaag accgagggca acatcaccga
     1081 gcaggtggag gacgccgtgg cccactcagc aggatcagga ggtggaggct gtgagcggga
     1141 gccccagagc ttgactcggg tggacctcag cagctcagtc ctcccaggag actctgtggg
     1201 gctggctgtc attctgcaca ctgacagcag gaaggactca ccaccagctg gaagtccagc
     1261 aggtagatca atctacaaca gcttttatgt gtattgcaaa ggcccctgtc aaagagtgca
     1321 gccgggaaaa ctcagggtac agtgcagcac ctgcaggcag gcaacgctca ccttgaccca
     1381 gggtccatct tgctgggatg atgttttaat tccaaaccgg atgagtggtg aatgccaatc
     1441 cccacactgc cctgggacta gtgcagaatt tttctttaaa tgtggagcac accccacctc
     1501 tgacaaggaa acatcagtag ctttgcacct gatcgcaaca aatagtcgga acatcacttg
     1561 cattacgtgc acagacgtca ggagccccgt cctggttttc cagtgcaact cccgccacgt
     1621 gatttgctta gactgtttcc acttatactg tgtgacaaga ctcaatgatc ggcagtttgt
     1681 ccacgaccct cagcttggct actccctgcc ttgtgtggct ggctgtccca actccttgat
     1741 taaagagctc catcacttca ggattctggg agaagagcag tacaaccggt accagcagta
     1801 tggtgcagag gagtgtgtcc tgcagatggg gggcgtgtta tgcccccgcc ctggctgtgg
     1861 agcggggctg ctgccggagc ctgaccagag gaaagtcacc tgcgaagggg gcaatggcct
     1921 gggctgtggg tttgccttct gccgggaatg taaagaagcg taccatgaag gggagtgcag
     1981 tgccgtattt gaagcctcag gaacaactac tcaggcctac agagtcgatg aaagagccgc
     2041 cgagcaggct cgttgggaag cagcctccaa agaaaccatc aagaaaacca ccaagccctg
     2101 tccccgctgc catgtaccag tggaaaaaaa tggaggctgc atgcacatga agtgtccgca
     2161 gccccagtgc aggctcgagt ggtgctggaa ctgtggctgc gagtggaacc gcgtctgcat
     2221 gggggaccac tggttcgacg tgtaaggatc caccgatgca tcgcgtctgg aacaatcaac
     2281 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2341 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2401 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2461 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2521 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2581 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2641 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     2701 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2761 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2821 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag
     2881 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     2941 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     3001 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     3061 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     3121 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
     3181 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
     3241 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
     3301 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
     3361 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
     3421 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     3481 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     3541 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     3601 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     3661 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     3721 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
     3781 attgctagcg tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5101 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5281 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6001 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6121 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
     6181 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
     6241 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
     6301 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta
     6361 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
     6421 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
     6481 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
     6541 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
     6601 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
     6661 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
     6721 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
     6781 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6841 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6901 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     6961 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     7021 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     7081 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     7141 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     7201 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     7261 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     7321 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     7381 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     7441 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     7501 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     7561 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     7621 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     7681 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     7741 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     7801 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     7861 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     7921 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     7981 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     8041 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     8101 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     8161 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     8221 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     8281 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg cctttaaaag
     8341 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     8401 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     8461 cagggac
//
