LOCUS       sequence-420271-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNAJC5 L116-Delta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  AUTHORS   Wu S, Hernandez Villegas NC, Sirkis DW, Thomas-Wright I, 
            Wade-Martins R, Schekman R
  TITLE     Unconventional secretion of alpha-synuclein mediated by 
            palmitoylated DNAJC5 oligomers.
  JOURNAL   Elife. 2023 Jan 10;12:e85837. doi: 10.7554/eLife.85837.
  PUBMED    36626307
REFERENCE   2  (bases 1 to 5447)
  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: "Elife."; 
            date: "2023-01-10"; volume: "12:e85837. doi"; pages: " 
            10.7554/eLife.85837"
COMMENT     Sequence Label: pCMV-DNAJC5-L116-Delta
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     610..629
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        610..628
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      656..665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1271..1300
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1319..1342
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1362..1378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1362..1378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1389..2011
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1588..1607)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2160..2748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2240..2259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3077..3124
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3149..3168
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3356..4150)
                     /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             complement(3356..4150)
                     /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(3467..3486)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4077..4096
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4185..4542)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4199..4334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4351..4542
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4499..4519
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4544..4648)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4675..5130
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4762..4781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4972..4993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5256..5278
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5270..5287
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5271..5287
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattttgt aatacgactc actatagggc ggccgggaat tcgtcgactg gatccgccac
      661 catggcagac cagagacagc gctcactgtc tacctctggg gagtcattgt accacgtcct
      721 tgggttggac aagaacgcaa cctcagatga cattaaaaag tcctatcgga agcttgcctt
      781 gaaatatcac cccgacaaga accccgacaa cccggaggcc gcggacaagt ttaaggagat
      841 caacaacgcg cacgccatcc tcacggacgc cacaaaaagg aacatctacg acaagtacgg
      901 ctcgctgggt ctctacgtgg ccgagcagtt tggggaagag aacgtgaaca cctacttcgt
      961 gctgtccagc tggtgggcca aggccctgtt tgtcttctgc ggcctcacgt gctgctactg
     1021 ctgctgctgt ctgtgctgct gcttcaactg ctgctgcggg aagtgtaagc ccaaggcgcc
     1081 tgaaggcgag gagacggagt tctacgtgtc ccccgaggat ctggaggcac agctgcagtc
     1141 tgacgagagg gaggccacag acacgccgat cgtcatacag ccggcatccg ccaccgagac
     1201 cacccagctc acagccgact cccaccccag ctaccacact gacgggttca acacgcgtac
     1261 gcggccgctc gagcagaaac tcatctcaga agaggatctg gcagcaaatg atatcctgga
     1321 ttacaaggat gacgacgata aggtttaaac ggccggccgc ggtcatagct gtttcctgaa
     1381 cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     1441 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     1501 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     1561 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     1621 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     1681 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     1741 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     1801 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     1861 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     1921 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     1981 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcggcg agcggtatca
     2041 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2581 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2821 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2881 tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg
     2941 gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg
     3001 cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
     3061 cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt
     3121 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
     3181 ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     3241 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     3301 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     3361 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     3421 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     3481 ccaacgctat gtcctgatag cgatccgcca cacccagccg gccacagtcg atgaatccag
     3541 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     3601 gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     3661 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     3721 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     3781 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     3841 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     3901 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     3961 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     4021 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     4081 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     4141 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
     4201 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     4261 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     4321 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     4381 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     4441 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4501 cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat
     4561 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4621 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     4681 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4741 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4801 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4861 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4921 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4981 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5041 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5101 attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg
     5161 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5221 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5281 ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat
     5341 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
     5401 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgtt
//
