LOCUS       sequence-420266-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNAJC5 (WT)-HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  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 6357)
  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 (WT)-HaloTag
FEATURES             Location/Qualifiers
     source          1..6357
                     /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"
     CDS             1287..2177
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             2181..2210
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2229..2252
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2272..2288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2272..2288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2299..2921
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2498..2517)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3070..3658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3150..3169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3987..4034
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4059..4078
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4266..5060)
                     /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(4266..5060)
                     /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(4377..4396)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4987..5006
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5095..5452)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5109..5244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5261..5452
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5409..5429
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5454..5558)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5585..6040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5672..5691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5882..5903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6166..6188
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6180..6197
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6181..6197
                     /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 gatccggtac
      661 cgaggagatc tgccgccgcg atcgccatgg cagaccagag acagcgctca ctgtctacct
      721 ctggggagtc attgtaccac gtccttgggt tggacaagaa cgcaacctca gatgacatta
      781 aaaagtccta tcggaagctt gccttgaaat atcaccccga caagaacccc gacaacccgg
      841 aggccgcgga caagtttaag gagatcaaca acgcgcacgc catcctcacg gacgccacaa
      901 aaaggaacat ctacgacaag tacggctcgc tgggtctcta cgtggccgag cagtttgggg
      961 aagagaacgt gaacacctac ttcgtgctgt ccagctggtg ggccaaggcc ctgtttgtct
     1021 tctgcggcct cctcacgtgc tgctactgct gctgctgtct gtgctgctgc ttcaactgct
     1081 gctgcgggaa gtgtaagccc aaggcgcctg aaggcgagga gacggagttc tacgtgtccc
     1141 ccgaggatct ggaggcacag ctgcagtctg acgagaggga ggccacagac acgccgatcg
     1201 tcatacagcc ggcatccgcc accgagacca cccagctcac agccgactcc caccccagct
     1261 accacactga cgggttcaac acgcgtatgg ctgaaatcgg cacagggttt cccttcgatc
     1321 ctcactatgt agaagtgctc ggggagcgaa tgcattatgt tgacgtaggg ccccgggacg
     1381 gtaccccagt actttttctt cacgggaacc ctacgtcctc atacgtgtgg cgaaacataa
     1441 taccacatgt ggctccgacg catagatgta tagcacctga cttgatcggg atgggcaaat
     1501 ccgacaagcc cgatctcgga tacttctttg acgatcatgt aaggttcatg gatgctttta
     1561 tcgaggcgtt gggccttgag gaggtagtat tggtgataca tgactgggga tctgcactgg
     1621 ggtttcattg ggcaaagcgc aatccggagc gcgtgaaagg cattgccttc atggagttta
     1681 tcagacccat ccctacctgg gacgagtggc cggagttcgc tagggagact tttcaagcgt
     1741 tccgcacaac tgacgtcgga cgcaagctca ttatagacca aaatgtcttt atcgaaggga
     1801 cactccccat gggagttgtg aggccgctga ctgaagttga gatggaccat tacagagaac
     1861 cattcctgaa ccccgtggat agagagccgc tttggcgctt cccaaatgag ctgcctattg
     1921 ctggcgaacc cgcaaatatc gtggccttgg ttgaggagta tatggattgg ctgcaccaga
     1981 gtccggtacc gaagctcctt ttctggggta cgccaggcgt cttgattcct ccggctgagg
     2041 ccgcgcgact tgctaaaagt cttcctaatt gcaaggcggt ggatattggc cccggcctga
     2101 acttgctcca ggaagataac ccggacctga ttggatcaga gatcgcccga tggttgtcta
     2161 ctctggaaat ctctggactc gagcagaaac tcatctcaga agaggatctg gcagcaaatg
     2221 atatcctgga ttacaaggat gacgacgata aggtttaaac ggccggccgc ggtcatagct
     2281 gtttcctgaa cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     2341 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     2401 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     2461 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     2521 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     2581 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     2641 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     2701 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     2761 cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag
     2821 cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc
     2881 ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcggcg
     2941 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3001 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3781 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc
     3841 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     3901 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     3961 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg
     4021 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt
     4081 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     4141 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     4201 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     4261 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     4321 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     4381 tcacgggtag ccaacgctat gtcctgatag cgatccgcca cacccagccg gccacagtcg
     4441 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     4501 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     4561 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     4621 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     4681 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     4741 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     4801 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     4861 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     4921 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     4981 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     5041 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     5101 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     5161 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     5221 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     5281 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     5341 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5401 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     5461 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5521 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5581 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5641 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5701 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5761 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5821 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5881 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5941 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6001 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     6061 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     6121 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6181 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
     6241 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     6301 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgtt
//
