LOCUS       sequence-420300-        6377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of XPACK-DNAJC5 (L115R)-HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6377)
  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 6377)
  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-XPACK-DNAJC5 (L115R)- HaloTag
FEATURES             Location/Qualifiers
     source          1..6377
                     /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             1307..2197
                     /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             2201..2230
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2249..2272
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2292..2308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2292..2308)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2319..2941
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2518..2537)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3090..3678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3170..3189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4007..4054
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4079..4098
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4286..5080)
                     /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(4286..5080)
                     /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(4397..4416)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5007..5026
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5115..5472)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5129..5264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5281..5472
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5429..5449
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5474..5578)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5605..6060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5692..5711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5902..5923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6186..6208
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6200..6217
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6201..6217
                     /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 catgggctgc attaacagca aacgcaaaga tgccggatcc accggtatgg cagaccagag
      721 acagcgctca ctgtctacct ctggggagtc attgtaccac gtccttgggt tggacaagaa
      781 cgcaacctca gatgacatta aaaagtccta tcggaagctt gccttgaaat atcaccccga
      841 caagaacccc gacaacccgg aggccgcgga caagtttaag gagatcaaca acgcgcacgc
      901 catcctcacg gacgccacaa aaaggaacat ctacgacaag tacggctcgc tgggtctcta
      961 cgtggccgag cagtttgggg aagagaacgt gaacacctac ttcgtgctgt ccagctggtg
     1021 ggccaaggcc ctgtttgtct tctgcggcag actcacgtgc tgctactgct gctgctgtct
     1081 gtgctgctgc ttcaactgct gctgcgggaa gtgtaagccc aaggcgcctg aaggcgagga
     1141 gacggagttc tacgtgtccc ccgaggatct ggaggcacag ctgcagtctg acgagaggga
     1201 ggccacagac acgccgatcg tcatacagcc ggcatccgcc accgagacca cccagctcac
     1261 agccgactcc caccccagct accacactga cgggttcaac acgcgtatgg ctgaaatcgg
     1321 cacagggttt cccttcgatc ctcactatgt agaagtgctc ggggagcgaa tgcattatgt
     1381 tgacgtaggg ccccgggacg gtaccccagt actttttctt cacgggaacc ctacgtcctc
     1441 atacgtgtgg cgaaacataa taccacatgt ggctccgacg catagatgta tagcacctga
     1501 cttgatcggg atgggcaaat ccgacaagcc cgatctcgga tacttctttg acgatcatgt
     1561 aaggttcatg gatgctttta tcgaggcgtt gggccttgag gaggtagtat tggtgataca
     1621 tgactgggga tctgcactgg ggtttcattg ggcaaagcgc aatccggagc gcgtgaaagg
     1681 cattgccttc atggagttta tcagacccat ccctacctgg gacgagtggc cggagttcgc
     1741 tagggagact tttcaagcgt tccgcacaac tgacgtcgga cgcaagctca ttatagacca
     1801 aaatgtcttt atcgaaggga cactccccat gggagttgtg aggccgctga ctgaagttga
     1861 gatggaccat tacagagaac cattcctgaa ccccgtggat agagagccgc tttggcgctt
     1921 cccaaatgag ctgcctattg ctggcgaacc cgcaaatatc gtggccttgg ttgaggagta
     1981 tatggattgg ctgcaccaga gtccggtacc gaagctcctt ttctggggta cgccaggcgt
     2041 cttgattcct ccggctgagg ccgcgcgact tgctaaaagt cttcctaatt gcaaggcggt
     2101 ggatattggc cccggcctga acttgctcca ggaagataac ccggacctga ttggatcaga
     2161 gatcgcccga tggttgtcta ctctggaaat ctctggactc gagcagaaac tcatctcaga
     2221 agaggatctg gcagcaaatg atatcctgga ttacaaggat gacgacgata aggtttaaac
     2281 ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc tgtgacccct
     2341 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     2401 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     2461 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     2521 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     2581 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2641 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2701 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     2761 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc
     2821 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga
     2881 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat
     2941 tgggtacgcg gccagcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3001 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3421 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3481 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3541 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3601 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3661 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3721 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3781 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag
     3841 gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
     3901 ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt
     3961 ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca
     4021 acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt
     4081 gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga
     4141 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     4201 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
     4261 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     4321 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     4381 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cgatccgcca
     4441 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     4501 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     4561 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     4621 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     4681 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     4741 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     4801 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     4861 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     4921 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     4981 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     5041 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     5101 tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     5161 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     5221 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     5281 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     5341 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     5401 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     5461 tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5521 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5581 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5641 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5701 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5761 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5821 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5881 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5941 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6001 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6061 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6121 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6181 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
     6241 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     6301 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
     6361 aatatgaccg ccatgtt
//
