LOCUS       sequence-420276-        5240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNAJC5-Delta-J (Delta-14-82).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5240)
  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 5240)
  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-J (Delta-14-82)
FEATURES             Location/Qualifiers
     source          1..5240
                     /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             1064..1093
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1112..1135
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1155..1171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1155..1171)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1182..1804
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1381..1400)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(1953..2541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2033..2052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2870..2917
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2942..2961
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3149..3943)
                     /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(3149..3943)
                     /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(3260..3279)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3870..3889
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3978..4335)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3992..4127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4144..4335
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4292..4312
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4337..4441)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4468..4923
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4555..4574)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4765..4786
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5049..5071
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5063..5080
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5064..5080
                     /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 tacctctggt ctctacgtgg ccgagcagtt
      721 tggggaagag aacgtgaaca cctacttcgt gctgtccagc tggtgggcca aggccctgtt
      781 tgtcttctgc ggcctcctca cgtgctgcta ctgctgctgc tgtctgtgct gctgcttcaa
      841 ctgctgctgc gggaagtgta agcccaaggc gcctgaaggc gaggagacgg agttctacgt
      901 gtcccccgag gatctggagg cacagctgca gtctgacgag agggaggcca cagacacgcc
      961 gatcgtcata cagccggcat ccgccaccga gaccacccag ctcacagccg actcccaccc
     1021 cagctaccac actgacgggt tcaacacgcg tacgcggccg ctcgagcaga aactcatctc
     1081 agaagaggat ctggcagcaa atgatatcct ggattacaag gatgacgacg ataaggttta
     1141 aacggccggc cgcggtcata gctgtttcct gaacagatcc cgggtggcat ccctgtgacc
     1201 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     1261 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     1321 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     1381 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     1441 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     1501 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     1561 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     1621 gattacaggc gtgaaccact gctcccttcc ctgtcctttt gattttaaaa taactatacc
     1681 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     1741 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     1801 aattgggtac gcggccagcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1861 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1921 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1981 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2041 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2101 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2161 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2221 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2281 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2341 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2401 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2461 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2521 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2581 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2641 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
     2701 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     2761 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     2821 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     2881 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     2941 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     3001 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     3061 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     3121 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     3181 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     3241 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcgatccg
     3301 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     3361 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     3421 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     3481 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     3541 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     3601 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     3661 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     3721 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     3781 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     3841 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     3901 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     3961 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     4021 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     4081 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     4141 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     4201 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     4261 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     4321 ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt
     4381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4441 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     4501 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4561 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4621 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4681 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4741 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4801 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4861 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4921 tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4981 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     5041 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt
     5101 gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg
     5161 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
     5221 tccaatatga ccgccatgtt
//
