LOCUS       sequence-424086-        5360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNAJC5-Delta-C30 (Delta-169-198).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5360)
  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 5360)
  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-Delta-C30 (Delta-169-198)
FEATURES             Location/Qualifiers
     source          1..5360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     230..249
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        230..248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      276..285
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             804..833
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             852..875
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(895..911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(895..911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    922..1544
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1121..1140)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(1693..2281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1773..1792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2610..2657
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2682..2701
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2889..3683)
                     /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(2889..3683)
                     /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(3000..3019)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3610..3629
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3718..4075)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3732..3867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3884..4075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4032..4052
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4077..4181)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4208..4663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4295..4314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4505..4526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4789..4811
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4803..4820
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4804..4820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4981..5360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc
      241 actatagggc ggccgggaat tcgtcgactg gatccgccac catggcagac cagagacagc
      301 gctcactgtc tacctctggg gagtcattgt accacgtcct tgggttggac aagaacgcaa
      361 cctcagatga cattaaaaag tcctatcgga agcttgcctt gaaatatcac cccgacaaga
      421 accccgacaa cccggaggcc gcggacaagt ttaaggagat caacaacgcg cacgccatcc
      481 tcacggacgc cacaaaaagg aacatctacg acaagtacgg ctcgctgggt ctctacgtgg
      541 ccgagcagtt tggggaagag aacgtgaaca cctacttcgt gctgtccagc tggtgggcca
      601 aggccctgtt tgtcttctgc ggcctcctca cgtgctgcta ctgctgctgc tgtctgtgct
      661 gctgcttcaa ctgctgctgc gggaagtgta agcccaaggc gcctgaaggc gaggagacgg
      721 agttctacgt gtcccccgag gatctggagg cacagctgca gtctgacgag agggaggcca
      781 cagacacgcg tacgcggccg ctcgagcaga aactcatctc agaagaggat ctggcagcaa
      841 atgatatcct ggattacaag gatgacgacg ataaggttta aacggccggc cgcggtcata
      901 gctgtttcct gaacagatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
      961 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     1021 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     1081 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     1141 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     1201 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     1261 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     1321 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     1381 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     1441 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     1501 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagcg
     1561 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2101 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2401 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     2461 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     2521 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     2581 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     2641 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     2701 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     2761 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     2821 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     2881 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     2941 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     3001 atatcacggg tagccaacgc tatgtcctga tagcgatccg ccacacccag ccggccacag
     3061 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     3121 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     3181 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     3241 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     3301 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     3361 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     3421 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     3481 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca
     3541 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     3601 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     3661 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt
     3721 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     3781 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     3841 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     3901 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     3961 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4021 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
     4081 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4141 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4201 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4261 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4321 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4381 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4441 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4561 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4621 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc
     4681 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     4741 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4801 gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat tggcaattag
     4861 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     4921 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
     4981 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     5041 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     5101 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     5161 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     5221 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     5281 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
     5341 tagtcatcgc tattaccatg
//
