LOCUS       sequence-494422-        5096 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 5096)
  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 5096)
  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 L115R
FEATURES             Location/Qualifiers
     source          1..5096
                     /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    2363..2985
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2562..2581)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3009..3029)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3014..3343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3194..3329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3420..3437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3591..4179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3671..3690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4350..5096,1..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     4973..4992
                     /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 tggccgcctc acctgctgct actgctgctg ctgtttatgc tgttgcttta
     2101 actgctgctg tgggaagtgc aagcccaagg cacctgaggg tgaagagaca gaattctacg
     2161 tatcccctga agacttggag gcacagctgc agtctgatga gagggaggct acagacacac
     2221 cgatcgtcat acagccagca tccgccacag agaccaccca gctgacagct gactcccacc
     2281 ccagctatca cactgacggg ttcaactaag tcgactctag attgcggccg cggtcatagc
     2341 tgtttcctcg agaacagatc ccgggtggca tccctgtgac ccctccccag tgcctctcct
     2401 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     2461 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     2521 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     2581 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     2641 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     2701 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     2761 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     2821 tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac
     2881 acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac
     2941 tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc
     3001 ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3061 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3181 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3301 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgactgcat
     3361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4021 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4261 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4321 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4381 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4441 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4501 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4561 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4621 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4681 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4741 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4801 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4861 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4921 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4981 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5041 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacc
//
