LOCUS       sequence-495963-        5295 bp    DNA     circular SYN 23-DEC-2025
DEFINITION  Mammalian expression of human DNAJC5 wildtype with a mCerulean tag 
            on its N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5295)
  AUTHORS   Lee J, Chin N, Zou J, Mazli WNAB, Jarnik M, Saidi L, Xu Y, Jeong E, 
            Suh J, Replogle J, Ward ME, Bonifacino JS, Zheng W, Hao L, Ye Y
  TITLE     CHIP protects lysosomes from CLN4 mutant-induced membrane damage.
  JOURNAL   Nat Cell Biol. 2025 Aug 25. doi: 10.1038/s41556-025-01738-2.
  PUBMED    40855364
REFERENCE   2  (bases 1 to 5295)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2025 Aug 25. doi: 10.1038/s41556-025-01738-2."
COMMENT     Sequence Label: mCerulean-DNAJC5
FEATURES             Location/Qualifiers
     source          1..5295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(100..688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(180..199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1017..1064
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1089..1108
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1296..2090)
                     /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(1296..2090)
                     /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(1407..1426)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2017..2036
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2125..2482)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2139..2274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2291..2482
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2439..2459
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2484..2588)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2615..3070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2702..2721)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2912..2933
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3077..3198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3088..3107)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3142..3161
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3952..4662,4663..4665,4666..4668))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3994..4015)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4343..4362
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4602..4623
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4665..4674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4713..4916)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4742..4762)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4917..5220
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
       61 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      481 gtatgtaggc ggtgatacag agttcttgaa gtggtggcct aactacggct acactagaag
      541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      781 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      841 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
      901 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
      961 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     1021 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     1081 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     1141 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     1201 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     1261 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     1321 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     1381 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     1441 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     1501 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     1561 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     1621 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     1681 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     1741 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     1801 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     1861 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     1921 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     1981 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     2041 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     2101 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     2161 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     2221 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     2281 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     2341 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     2401 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     2461 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
     2521 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2581 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     2641 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2701 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2761 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2821 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2881 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2941 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3001 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3061 cttacaattt acgccttaag atacattgat gagtttggac aaaccacaac tagaatgcag
     3121 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3181 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3241 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     3301 gatcagttat ctagatccgg tggatcctta gttgaacccg tcggtgtgat agctggggtg
     3361 ggagtcagct gtcagctggg tggtctctgt ggcggatgct ggctgtatga cgatcggtgt
     3421 gtctgtagcc tccctctcat cagactgcag ctgtgcctcc aagtcttcag gggatacgta
     3481 gaattctgtc tcttcaccct caggtgcctt gggcttgcac ttcccacagc agcagttaaa
     3541 gcaacagcat aaacagcagc agcagtagca gcaggtgagg aggccacaga agacgaacag
     3601 cgccttggcc caccagctgg agagtacgaa gtaggtgttg acattctcct ccccaaattg
     3661 ctcagccaca tagagcccca gcgagccata cttgtcataa atgtttcttt tcgtggcgtc
     3721 tgtcaggatt gcgtgggcgt tgttaatctc cttaaacttg tctgcagcct ctgggttatc
     3781 agggttcttg tcagggtgat atttcagggc cagcttccga taggactttt taatgtcatc
     3841 tgaggttgca ttcttgtcca gtccaagaac atggtataac gattccccgg aagtagagag
     3901 tgagcgctgc ctctggtcag ccgaagcttg agctcgagat ctgagtccgg acttgtacag
     3961 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     4021 gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg
     4081 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     4141 gctgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt
     4201 gtcggcggtg atatagacgt tgtcgctgat ggcgttgtac tccagcttgt gccccaggat
     4261 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     4321 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     4381 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     4441 ggggtagcgg gcgaagcact gcacgcccca ggtcagggtg gtcacgaggg tgggccaggg
     4501 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     4561 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     4621 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg
     4681 tagcgctagc ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt
     4741 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     4801 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     4861 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     4921 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     4981 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     5041 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     5101 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     5161 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     5221 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     5281 atgcatggcg gtaat
//
