LOCUS       sequence-420270-        5390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNAJC5-Delta-C20 (Delta-179-198).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5390)
  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 5390)
  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-C20 (Delta-179-198)
FEATURES             Location/Qualifiers
     source          1..5390
                     /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             1214..1243
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1262..1285
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1305..1321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1305..1321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1332..1954
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1531..1550)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2103..2691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2183..2202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3020..3067
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3092..3111
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3299..4093)
                     /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(3299..4093)
                     /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(3410..3429)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4020..4039
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4128..4485)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4142..4277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4294..4485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4442..4462
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4487..4591)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4618..5073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4705..4724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4915..4936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5199..5221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5213..5230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5214..5230
                     /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 ggcctcctca cgtgctgcta
     1021 ctgctgctgc tgtctgtgct gctgcttcaa ctgctgctgc gggaagtgta agcccaaggc
     1081 gcctgaaggc gaggagacgg agttctacgt gtcccccgag gatctggagg cacagctgca
     1141 gtctgacgag agggaggcca cagacacgcc gatcgtcata cagccggcat ccgccacgcg
     1201 tacgcggccg ctcgagcaga aactcatctc agaagaggat ctggcagcaa atgatatcct
     1261 ggattacaag gatgacgacg ataaggttta aacggccggc cgcggtcata gctgtttcct
     1321 gaacagatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     1381 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     1441 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     1501 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     1561 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     1621 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     1681 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     1741 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     1801 ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc
     1861 tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat
     1921 gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagcg gcgagcggta
     1981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2281 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2521 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2821 aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg
     2881 ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa
     2941 acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     3001 gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt
     3061 attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc
     3121 cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     3181 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     3241 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     3301 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     3361 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     3421 tagccaacgc tatgtcctga tagcgatccg ccacacccag ccggccacag tcgatgaatc
     3481 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     3541 cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga
     3601 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     3661 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     3721 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     3781 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     3841 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     3901 ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     3961 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     4021 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     4081 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct
     4141 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg
     4201 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     4261 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     4321 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     4381 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     4441 caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc
     4501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4621 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4681 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4741 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4801 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4861 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4921 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4981 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5041 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
     5101 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5161 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5221 gacggccagt gccaagctga tctatacatt gaatcaatat tggcaattag ccatattagt
     5281 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
     5341 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
//
