LOCUS       sequence-420267-        6332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNAJC5 (L115R)-HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6332)
  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 6332)
  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 (L115R)-HaloTag
FEATURES             Location/Qualifiers
     source          1..6332
                     /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             1262..2152
                     /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             2156..2185
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2204..2227
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2247..2263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2247..2263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2274..2896
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2473..2492)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3045..3633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3125..3144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3962..4009
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4034..4053
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4241..5035)
                     /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(4241..5035)
                     /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(4352..4371)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4962..4981
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5070..5427)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5084..5219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5236..5427
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5384..5404
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5429..5533)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5560..6015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5647..5666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5857..5878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6141..6163
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6155..6172
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6156..6172
                     /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 ggcagactca cgtgctgcta
     1021 ctgctgctgc tgtctgtgct gctgcttcaa ctgctgctgc gggaagtgta agcccaaggc
     1081 gcctgaaggc gaggagacgg agttctacgt gtcccccgag gatctggagg cacagctgca
     1141 gtctgacgag agggaggcca cagacacgcc gatcgtcata cagccggcat ccgccaccga
     1201 gaccacccag ctcacagccg actcccaccc cagctaccac actgacgggt tcaacacgcg
     1261 tatggctgaa atcggcacag ggtttccctt cgatcctcac tatgtagaag tgctcgggga
     1321 gcgaatgcat tatgttgacg tagggccccg ggacggtacc ccagtacttt ttcttcacgg
     1381 gaaccctacg tcctcatacg tgtggcgaaa cataatacca catgtggctc cgacgcatag
     1441 atgtatagca cctgacttga tcgggatggg caaatccgac aagcccgatc tcggatactt
     1501 ctttgacgat catgtaaggt tcatggatgc ttttatcgag gcgttgggcc ttgaggaggt
     1561 agtattggtg atacatgact ggggatctgc actggggttt cattgggcaa agcgcaatcc
     1621 ggagcgcgtg aaaggcattg ccttcatgga gtttatcaga cccatcccta cctgggacga
     1681 gtggccggag ttcgctaggg agacttttca agcgttccgc acaactgacg tcggacgcaa
     1741 gctcattata gaccaaaatg tctttatcga agggacactc cccatgggag ttgtgaggcc
     1801 gctgactgaa gttgagatgg accattacag agaaccattc ctgaaccccg tggatagaga
     1861 gccgctttgg cgcttcccaa atgagctgcc tattgctggc gaacccgcaa atatcgtggc
     1921 cttggttgag gagtatatgg attggctgca ccagagtccg gtaccgaagc tccttttctg
     1981 gggtacgcca ggcgtcttga ttcctccggc tgaggccgcg cgacttgcta aaagtcttcc
     2041 taattgcaag gcggtggata ttggccccgg cctgaacttg ctccaggaag ataacccgga
     2101 cctgattgga tcagagatcg cccgatggtt gtctactctg gaaatctctg gactcgagca
     2161 gaaactcatc tcagaagagg atctggcagc aaatgatatc ctggattaca aggatgacga
     2221 cgataaggtt taaacggccg gccgcggtca tagctgtttc ctgaacagat cccgggtggc
     2281 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     2341 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     2401 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     2461 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     2521 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     2581 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     2641 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     2701 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa
     2761 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac
     2821 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt
     2881 agatgcctgt tgaattgggt acgcggccag cggcgagcgg tatcagctca ctcaaaggcg
     2941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3781 tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg
     3841 ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc
     3901 caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta
     3961 tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
     4021 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag
     4081 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
     4141 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
     4201 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     4261 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     4321 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     4381 gatagcgatc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     4441 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     4501 gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     4561 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     4621 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     4681 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     4741 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     4801 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt
     4861 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     4921 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     4981 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     5041 acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct
     5101 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     5161 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     5221 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     5281 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     5341 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     5401 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt
     5461 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5521 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     5581 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5641 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5701 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5761 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5821 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5881 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5941 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6001 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6061 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6121 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     6181 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
     6241 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
     6301 tattggctca tgtccaatat gaccgccatg tt
//
