LOCUS       sequence-495980-        5295 bp    DNA     circular SYN 23-DEC-2025
DEFINITION  Mammalian expression of human DNAJC5 L115R with a mCitrine (YFP) 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: mCitrine-DNAJC5 L115R
FEATURES             Location/Qualifiers
     source          1..5295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..110)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    135..182
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      511..1099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1000..1019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1274..1577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1578..1781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1732..1752
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1820..1829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1826..1828,1829..1831,1832..2542)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1871..1892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2132..2151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2479..2500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3296..3417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3333..3352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3387..3406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3424..3879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3773..3792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3906..4010
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4012..4369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4012..4203
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4035..4055)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4220..4355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4404..5198
                     /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             4404..5198
                     /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(4458..4477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5068..5087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
       61 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
      121 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
      181 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
      241 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
      301 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
      361 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
      421 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
      481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1141 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1201 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     1261 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     1321 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     1381 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     1441 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     1501 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     1561 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     1621 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     1681 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     1741 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     1801 tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     1861 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1921 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1981 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt
     2041 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2101 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2161 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2221 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     2281 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     2341 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2401 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     2461 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2521 cggcatggac gagctgtaca agtccggact cagatctcga gctcaagctt cggctgacca
     2581 gaggcagcgc tcactctcta cttccgggga atcgttatac catgttcttg gactggacaa
     2641 gaatgcaacc tcagatgaca ttaaaaagtc ctatcggaag ctggccctga aatatcaccc
     2701 tgacaagaac cctgataacc cagaggctgc agacaagttt aaggagatta acaacgccca
     2761 cgcaatcctg acagacgcca cgaaaagaaa catttatgac aagtatggct cgctggggct
     2821 ctatgtggct gagcaatttg gggaggagaa tgtcaacacc tacttcgtac tctccagctg
     2881 gtgggccaag gcgctgttcg tcttctgtgg ccgcctcacc tgctgctact gctgctgctg
     2941 tttatgctgt tgctttaact gctgctgtgg gaagtgcaag cccaaggcac ctgagggtga
     3001 agagacagaa ttctacgtat cccctgaaga cttggaggca cagctgcagt ctgatgagag
     3061 ggaggctaca gacacaccga tcgtcataca gccagcatcc gccacagaga ccacccagct
     3121 gacagctgac tcccacccca gctatcacac cgacgggttc aactaaggat ccaccggatc
     3181 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3241 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3301 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3361 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     3421 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3481 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3541 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3601 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3661 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3721 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3781 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3841 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3901 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3961 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4021 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4081 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4141 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4201 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4261 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4321 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4381 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4441 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4501 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4561 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4621 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4681 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4741 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4801 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4861 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4921 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4981 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5041 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5101 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5161 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5221 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5281 tgaaaggttg ggctt
//
