LOCUS       sequence-391244-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of untagged Parkin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5351)
  AUTHORS   Harding O, Holzer E, Riley JF, Martens S, Holzbaur ELF
  TITLE     Damaged mitochondria recruit the effector NEMO to activate NF-kappaB
            signaling.
  JOURNAL   Mol Cell. 2023 Sep 7;83(17):3188-3204.e7. doi: 
            10.1016/j.molcel.2023.08.005.
  PUBMED    37683611
REFERENCE   2  (bases 1 to 5351)
  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"; doi: 
            "10.1016/j.molcel.2023.08"; journalName: "Mol Cell"; date: 
            "2023-09-7- 7"; volume: "83"; issue: "17"; pages: "3188-3204.e7"
COMMENT     Sequence Label: Parkin
FEATURES             Location/Qualifiers
     source          1..5351
                     /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"
     polyA_signal    2139..2260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2176..2195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2230..2249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2267..2722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2404..2425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2616..2635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2749..2853
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2855..3212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2855..3046
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2878..2898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3063..3198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3247..4041
                     /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             3247..4041
                     /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(3301..3320)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3911..3930
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4229..4248)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4273..4320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4649..5237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5138..5157
                     /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 ccggtgccac catgatagtg tttgtcaggt tcaactccag ccatggtttc ccagtggagg
      661 tcgattctga caccagcatc ttccagctca aggaggtggt tgctaagcga cagggggttc
      721 cggctgacca gttgcgtgtg attttcgcag ggaaggagct gaggaatgac tggactgtgc
      781 agaattgtga cctggatcag cagagcattg ttcacattgt gcagagaccg tggagaaaag
      841 gtcaagaaat gaatgcaact ggaggcgacg accccagaaa cgcggcggga ggctgtgagc
      901 gggagcccca gagcttgact cgggtggacc tcagcagctc agtcctccca ggagactctg
      961 tggggctggc tgtcattctg cacactgaca gcaggaagga ctcaccacca gctggaagtc
     1021 cagcaggtag atcaatctac aacagctttt atgtgtattg caaaggcccc tgtcaaagag
     1081 tgcagccggg aaaactcagg gtacagtgca gcacctgcag gcaggcaacg ctcaccttga
     1141 cccagggtcc atcttgctgg gatgatgttt taattccaaa ccggatgagt ggtgaatgcc
     1201 aatccccaca ctgccctggg actagtgcag aatttttctt taaatgtgga gcacacccca
     1261 cctctgacaa ggaaacatca gtagctttgc acctgatcgc aacaaatagt cggaacatca
     1321 cttgcattac gtgcacagac gtcaggagcc ccgtcctggt tttccagtgc aactcccgcc
     1381 acgtgatttg cttagactgt ttccacttat actgtgtgac aagactcaat gatcggcagt
     1441 ttgtccacga ccctcagctt ggctactccc tgccttgtgt ggctggctgt cccaactcct
     1501 tgattaaaga gctccatcac ttcaggattc tgggagaaga gcagtacaac cggtaccagc
     1561 agtatggtgc agaggagtgt gtcctgcaga tggggggcgt gttatgcccc cgccctggct
     1621 gtggagcggg gctgctgccg gagcctgacc agaggaaagt cacctgcgaa gggggcaatg
     1681 gcctgggctg tgggtttgcc ttctgccggg aatgtaaaga agcgtaccat gaaggggagt
     1741 gcagtgccgt atttgaagcc tcaggaacaa ctactcaggc ctacagagtc gatgaaagag
     1801 ccgccgagca ggctcgttgg gaagcagcct ccaaagaaac catcaagaaa accaccaagc
     1861 cctgtccccg ctgccatgta ccagtggaaa aaaatggagg ctgcatgcac atgaagtgtc
     1921 cgcagcccca gtgcaggctc gagtggtgct ggaactgtgg ctgcgagtgg aaccgcgtct
     1981 gcatggggga ccactggttc gacgtgtaag gatccaccgg atctagataa ctgatcataa
     2041 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2101 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2161 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2221 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2281 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2341 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2401 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2461 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2521 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2581 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2641 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2701 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2761 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2821 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2881 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3121 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3181 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3241 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3301 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3361 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3421 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3481 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3541 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3601 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3661 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3721 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3781 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3841 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3901 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3961 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4021 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4081 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4141 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4201 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4261 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4321 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4381 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4441 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4501 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4561 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4621 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4681 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4741 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4801 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4861 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4921 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4981 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5041 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5101 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5161 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5221 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5281 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5341 ccgccatgca t
//
