LOCUS       sequence-421787-        5352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient mammalian expresssion of E16A 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(E16A)
FEATURES             Location/Qualifiers
     source          1..5352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        28..331
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        332..535
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     486..506
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2107..2228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2144..2163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2198..2217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2235..2690)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2372..2393)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2584..2603
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2717..2821
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2823..3180
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2823..3014
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2846..2866)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3031..3166
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3215..4009
                     /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             3215..4009
                     /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(3269..3288)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3879..3898
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4197..4216)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4241..4288
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4617..5205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5106..5125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
       61 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      121 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      181 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      241 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      301 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      361 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      421 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      481 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      541 agtgaaccgt cagatccgct agcgctaccg gtcgccacca tgatagtgtt tgtcaggttc
      601 aactccagcc atggtttccc agtggcggtc gattctgaca ccagcatctt ccagctcaag
      661 gaggtggttg ctaagcgaca gggggttccg gctgaccagt tgcgtgtgat tttcgcaggg
      721 aaggagctga ggaatgactg gactgtgcag aattgtgacc tggatcagca gagcattgtt
      781 cacattgtgc agagaccgtg gagaaaaggt caagaaatga atgcaactgg aggcgacgac
      841 cccagaaacg cggcgggagg ctgtgagcgg gagccccaga gcttgactcg ggtggacctc
      901 agcagctcag tcctcccagg agactctgtg gggctggctg tcattctgca cactgacagc
      961 aggaaggact caccaccagc tggaagtcca gcaggtagat caatctacaa cagcttttat
     1021 gtgtattgca aaggcccctg tcaaagagtg cagccgggaa aactcagggt acagtgcagc
     1081 acctgcaggc aggcaacgct caccttgacc cagggtccat cttgctggga tgatgtttta
     1141 attccaaacc ggatgagtgg tgaatgccaa tccccacact gccctgggac tagtgcagaa
     1201 tttttcttta aatgtggagc acaccccacc tctgacaagg aaacatcagt agctttgcac
     1261 ctgatcgcaa caaatagtcg gaacatcact tgcattacgt gcacagacgt caggagcccc
     1321 gtcctggttt tccagtgcaa ctcccgccac gtgatttgct tagactgttt ccacttatac
     1381 tgtgtgacaa gactcaatga tcggcagttt gtccacgacc ctcagcttgg ctactccctg
     1441 ccttgtgtgg ctggctgtcc caactccttg attaaagagc tccatcactt caggattctg
     1501 ggagaagagc agtacaaccg gtaccagcag tatggtgcag aggagtgtgt cctgcagatg
     1561 gggggcgtgt tatgcccccg ccctggctgt ggagcggggc tgctgccgga gcctgaccag
     1621 aggaaagtca cctgcgaagg gggcaatggc ctgggctgtg ggtttgcctt ctgccgggaa
     1681 tgtaaagaag cgtaccatga aggggagtgc agtgccgtat ttgaagcctc aggaacaact
     1741 actcaggcct acagagtcga tgaaagagcc gccgagcagg ctcgttggga agcagcctcc
     1801 aaagaaacca tcaagaaaac caccaagccc tgtccccgct gccatgtacc agtggaaaaa
     1861 aatggaggct gcatgcacat gaagtgtccg cagccccagt gcaggctcga gtggtgctgg
     1921 aactgtggct gcgagtggaa ccgcgtctgc atgggggacc actggttcga cgtgtaagga
     1981 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2041 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2101 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2161 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2221 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2281 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2341 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2401 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2461 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2521 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2581 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2641 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     2701 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2761 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2821 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2881 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2941 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3001 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3061 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3121 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3181 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3241 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3301 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3361 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3421 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3481 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3541 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3601 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3661 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3721 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3781 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3841 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3901 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     3961 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4021 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4081 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4141 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4201 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4261 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4321 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4381 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4441 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4741 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5281 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     5341 tacggggtca tt
//
