LOCUS       sequence-421788-        5352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient mammalian expresssion of untagged Parkin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5352)
  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 5352)
  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: pCMV-Parkin
FEATURES             Location/Qualifiers
     source          1..5352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        36..339
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        340..543
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     494..514
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2115..2236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2152..2171)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2206..2225
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2243..2698)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2380..2401)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2592..2611
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2725..2829
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2831..3188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2831..3022
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2854..2874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3039..3174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3223..4017
                     /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             3223..4017
                     /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(3277..3296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3887..3906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4205..4224)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4249..4296
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4625..5213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5114..5133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
       61 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      121 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      181 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      241 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      301 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      361 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      421 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      481 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      541 gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg atagtgtttg
      601 tcaggttcaa ctccagccat ggtttcccag tggaggtcga ttctgacacc agcatcttcc
      661 agctcaagga ggtggttgct aagcgacagg gggttccggc tgaccagttg cgtgtgattt
      721 tcgcagggaa ggagctgagg aatgactgga ctgtgcagaa ttgtgacctg gatcagcaga
      781 gcattgttca cattgtgcag agaccgtgga gaaaaggtca agaaatgaat gcaactggag
      841 gcgacgaccc cagaaacgcg gcgggaggct gtgagcggga gccccagagc ttgactcggg
      901 tggacctcag cagctcagtc ctcccaggag actctgtggg gctggctgtc attctgcaca
      961 ctgacagcag gaaggactca ccaccagctg gaagtccagc aggtagatca atctacaaca
     1021 gcttttatgt gtattgcaaa ggcccctgtc aaagagtgca gccgggaaaa ctcagggtac
     1081 agtgcagcac ctgcaggcag gcaacgctca ccttgaccca gggtccatct tgctgggatg
     1141 atgttttaat tccaaaccgg atgagtggtg aatgccaatc cccacactgc cctgggacta
     1201 gtgcagaatt tttctttaaa tgtggagcac accccacctc tgacaaggaa acatcagtag
     1261 ctttgcacct gatcgcaaca aatagtcgga acatcacttg cattacgtgc acagacgtca
     1321 ggagccccgt cctggttttc cagtgcaact cccgccacgt gatttgctta gactgtttcc
     1381 acttatactg tgtgacaaga ctcaatgatc ggcagtttgt ccacgaccct cagcttggct
     1441 actccctgcc ttgtgtggct ggctgtccca actccttgat taaagagctc catcacttca
     1501 ggattctggg agaagagcag tacaaccggt accagcagta tggtgcagag gagtgtgtcc
     1561 tgcagatggg gggcgtgtta tgcccccgcc ctggctgtgg agcggggctg ctgccggagc
     1621 ctgaccagag gaaagtcacc tgcgaagggg gcaatggcct gggctgtggg tttgccttct
     1681 gccgggaatg taaagaagcg taccatgaag gggagtgcag tgccgtattt gaagcctcag
     1741 gaacaactac tcaggcctac agagtcgatg aaagagccgc cgagcaggct cgttgggaag
     1801 cagcctccaa agaaaccatc aagaaaacca ccaagccctg tccccgctgc catgtaccag
     1861 tggaaaaaaa tggaggctgc atgcacatga agtgtccgca gccccagtgc aggctcgagt
     1921 ggtgctggaa ctgtggctgc gagtggaacc gcgtctgcat gggggaccac tggttcgacg
     1981 tgtaaggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2041 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2101 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2161 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2221 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2281 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2341 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2401 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2461 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2521 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2581 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2641 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2701 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2761 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2821 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2881 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2941 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3001 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3061 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3121 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3181 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3241 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3301 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3361 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3421 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3481 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3541 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3601 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3661 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3721 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     3781 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3841 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3901 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3961 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4021 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4081 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4141 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4201 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4261 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4321 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4381 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4441 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4501 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4561 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4621 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4681 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4741 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4801 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4861 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4921 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4981 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5041 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5101 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5161 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5221 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5281 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     5341 taatcaatta cg
//
