LOCUS       sequence-494416-        5093 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  Mammalian expression of human DNAJC5 with L115R mutation, and a FLAG
            tag on its N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5093)
  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 5093)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 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: pCMV6 FLAG-DNAJC5 delL116
FEATURES             Location/Qualifiers
     source          1..5093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(115..219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      246..701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(333..352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     543..564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     827..849
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     841..858
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     842..858
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        1019..1398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1399..1602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1553..1573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1604..1622
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1604..1621
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1656..1665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1665..1688
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2360..2982
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2559..2578)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3006..3026)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3011..3340
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3191..3326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3417..3434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3588..4176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3668..3687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4347..5093,1..45,46..114))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4970..4989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
       61 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      241 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      301 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      361 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      421 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
      481 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      541 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      601 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      661 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
      721 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      781 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      841 tgtaaaacga cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc
      901 atattagtca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
      961 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttga
     1021 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     1081 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1141 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1201 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1261 gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1321 cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta
     1381 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg
     1441 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1501 caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg
     1561 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcatttagg tgacactata
     1621 gaatacaagc tacttgttct ttttgcagcg gcttcgccac catggactac aaagacgatg
     1681 acgacaaggg atccgaattc agatctggta ccatggcaga ccagagacag cgctcactct
     1741 ctacttccgg ggaatcgtta taccatgttc ttggactgga caagaatgca acctcagatg
     1801 acattaaaaa gtcctatcgg aagctggccc tgaaatatca ccctgacaag aaccctgata
     1861 acccagaggc tgcagacaag tttaaggaga ttaacaacgc ccacgcaatc ctgacagacg
     1921 ccacgaaaag aaacatttat gacaagtatg gctcgctggg gctctatgtg gctgagcaat
     1981 ttggggagga gaatgtcaac acctacttcg tactctccag ctggtgggcc aaggcgctgt
     2041 tcgtcttctg tggcctcacc tgctgctact gctgctgctg tttatgctgt tgctttaact
     2101 gctgctgtgg gaagtgcaag cccaaggcac ctgagggtga agagacagaa ttctacgtat
     2161 cccctgaaga cttggaggca cagctgcagt ctgatgagag ggaggctaca gacacaccga
     2221 tcgtcataca gccagcatcc gccacagaga ccacccagct gacagctgac tcccacccca
     2281 gctatcacac tgacgggttc aactaagtcg actctagatt gcggccgcgg tcatagctgt
     2341 ttcctcgaga acagatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     2401 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     2461 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     2521 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     2581 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2641 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2701 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     2761 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     2821 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
     2881 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt
     2941 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg
     3001 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3061 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3181 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3241 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3301 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg actgcattaa
     3361 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3421 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3481 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3541 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3601 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3661 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3721 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3781 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3841 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3901 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3961 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4021 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4081 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4141 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4201 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4261 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4321 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4381 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4441 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4501 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4561 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4621 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4681 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4741 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4801 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4861 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4921 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4981 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5041 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acc
//
