LOCUS       sequence-420273-        5450 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of DNAJC5 L115R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5450)
  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 5450)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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
FEATURES             Location/Qualifiers
     source          1..5450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     230..249
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        230..248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      276..285
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             894..923
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             942..965
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(985..1001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(985..1001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1012..1634
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1211..1230)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(1783..2371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1863..1882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2700..2747
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2772..2791
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2979..3773)
                     /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(2979..3773)
                     /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(3090..3109)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3700..3719
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3808..4165)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3822..3957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3974..4165
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4122..4142
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4167..4271)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4298..4753
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4385..4404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4595..4616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4879..4901
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4893..4910
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4894..4910
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5071..5450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc
      241 actatagggc ggccgggaat tcgtcgactg gatccgccac catggcagac cagagacagc
      301 gctcactgtc tacctctggg gagtcattgt accacgtcct tgggttggac aagaacgcaa
      361 cctcagatga cattaaaaag tcctatcgga agcttgcctt gaaatatcac cccgacaaga
      421 accccgacaa cccggaggcc gcggacaagt ttaaggagat caacaacgcg cacgccatcc
      481 tcacggacgc cacaaaaagg aacatctacg acaagtacgg ctcgctgggt ctctacgtgg
      541 ccgagcagtt tggggaagag aacgtgaaca cctacttcgt gctgtccagc tggtgggcca
      601 aggccctgtt tgtcttctgc ggcagactca cgtgctgcta ctgctgctgc tgtctgtgct
      661 gctgcttcaa ctgctgctgc gggaagtgta agcccaaggc gcctgaaggc gaggagacgg
      721 agttctacgt gtcccccgag gatctggagg cacagctgca gtctgacgag agggaggcca
      781 cagacacgcc gatcgtcata cagccggcat ccgccaccga gaccacccag ctcacagccg
      841 actcccaccc cagctaccac actgacgggt tcaacacgcg tacgcggccg ctcgagcaga
      901 aactcatctc agaagaggat ctggcagcaa atgatatcct ggattacaag gatgacgacg
      961 ataaggttta aacggccggc cgcggtcata gctgtttcct gaacagatcc cgggtggcat
     1021 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     1081 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     1141 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     1201 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     1261 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     1321 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     1381 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     1441 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa
     1501 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg
     1561 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag
     1621 atgcctgttg aattgggtac gcggccagcg gcgagcggta tcagctcact caaaggcggt
     1681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2521 tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc
     2581 ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca
     2641 atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg
     2701 aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     2761 tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa
     2821 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     2881 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     2941 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     3001 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     3061 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     3121 tagcgatccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     3181 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     3241 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     3301 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     3361 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     3421 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     3481 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     3541 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     3601 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     3661 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     3721 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     3781 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc
     3841 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     3901 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     3961 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     4021 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     4081 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     4141 ccacagctgg ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat
     4201 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4261 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     4321 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4381 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4441 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4501 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4561 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4621 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4681 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4741 acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4801 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     4861 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctga
     4921 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     4981 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     5041 ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt
     5101 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     5161 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     5221 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     5281 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
     5341 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
     5401 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
