LOCUS       sequence-472711-        8709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the C-terminal fragments of VPS13C in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8709)
  AUTHORS   Wang X, Xu P, Bentley-DeSousa A, Hancock-Cerutti W, Cai S, Johnson 
            BT, Tonelli F, Shao L, Talaia G, Alessi DR, Ferguson SM, De Camilli 
            P
  TITLE     The bridge-like lipid transport protein VPS13C/PARK23 mediates 
            ER-lysosome contacts following lysosome damage.
  JOURNAL   Nat Cell Biol. 2025 May;27(5):776-789. doi: 
            10.1038/s41556-025-01653-6. Epub 2025 Apr 10.
  PUBMED    40211074
REFERENCE   2  (bases 1 to 8709)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41556-025-01653-6"; journalName: "Nat Cell Biol"; date: 
            "2025-05"; volume: "27"; issue: "5"; pages: "776-789"
COMMENT     Sequence Label: mCherry-VPS13C-Cter
FEATURES             Location/Qualifiers
     source          1..8709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      371..380
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(393..1100)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(668..687)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     937..955
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1097..1106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1144..1347)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1173..1193)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1348..1651
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1826..2414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1906..1925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2743..2790
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2815..2834
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3022..3816)
                     /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(3022..3816)
                     /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(3133..3152)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3743..3762
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3851..4208)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3865..4000
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4017..4208
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4165..4185
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4210..4314)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4341..4796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4638..4659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4803..4924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4814..4833)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4868..4887
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 actttattcc cttcagttgc atcaatttgt accaagacag agtcagaatg actggcattg
       61 ggattccgta cattatacaa tcgccgtcca ggtctggcca cagggatatt tgcaacttct
      121 gtatatccat gaggtacaat ggtcagagtg aagaaggagc tttcttgacg gctcaatata
      181 gataggttcc cttgacttga aggtaccatg ctggcatact ccagttccaa attctggcca
      241 gcatcaacat caaaaatatc acttttctca gggaagccca ttactctgag attacaattg
      301 ggcttcacct taatgggaac acctacagca ttttttaccg taaaaggagc tctgtccttt
      361 aaagagtagt ccatggtggc atctcgagat ctcttgtaca gctcgtccat gccgccggtg
      421 gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag
      481 gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc
      541 ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc
      601 ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag
      661 cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc
      721 accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg
      781 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag
      841 tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg
      901 gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc
      961 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
     1021 acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc
     1081 tcctcgccct tgctcaccat ggtggcaccg gtagcgctag cggatctgac ggttcactaa
     1141 accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     1201 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     1261 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     1321 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     1381 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     1441 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     1501 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     1561 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     1621 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     1681 tgaccccgta attgattact attaataact aatgcatggc ggtaatacgg ttatccacag
     1741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta
     2581 tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg
     2641 ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg
     2701 tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac
     2761 cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct
     2821 ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc
     2881 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
     2941 agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca
     3001 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     3061 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     3121 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     3181 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     3241 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct
     3301 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     3361 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     3421 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     3481 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     3541 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     3601 cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag
     3661 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     3721 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     3781 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     3841 ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     3901 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     3961 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     4021 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     4081 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     4141 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
     4201 cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4261 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4321 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4681 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc
     4801 cttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4861 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4921 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4981 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agttatctag
     5041 atccggtgga tcccgggccc ttaagatggc aattggggtc tgagaagtct cactgatgac
     5101 tgcttcatca atttttgctg ctgtctcgtt gactgtgcat cctcaatggc attacatgct
     5161 ctctctgctg tggcggtgtc cttcaggtaa acttttcgaa cacagccttg attggcactg
     5221 tcttttttgt ggaacagacc ctgttcctta actgaaattt ttagcacatt ttcactgaca
     5281 ctaggaggaa atacaaaatc ttcaaatgga cattgccagt ctacacacat aaggcccagg
     5341 atttcaactt cctttataca caacactcgc ctatttgtaa ccataaggat tgtcttcttg
     5401 cttccaggaa tagcacagtg gtatcggtaa gtctctcctt ccaacttttt gatatgattc
     5461 tcaagtaagt cagagccctc agattcctgt ctgtcataag gacgaatgat gccatcttca
     5521 tggatcaggc gagggggacg gaggctagat acttcctcag ttgattctgc tgccctctga
     5581 atgccttgga aggtactact ggccatatct acgattccac cagttggacg ggccacagca
     5641 cccacaagcc cttttccaat tcctttaaag aatccagcag ctccttcctt tttggcacct
     5701 tccacaggtt ttgttattat tccagtcact ccaccaacaa ctcctcgcag aaagcccttt
     5761 cctcctctgg ccaggctgtc tccaaaatct ctgggctgtc gactcaactc ttctcttctt
     5821 ttttgctgat attccttgtc cattgtaatt gctgccaaac ctttcccaac agaaccggtg
     5881 attcgagata caactcctgc tgcaccacct actgtgtgtc caaagaggct tctcactcca
     5941 atcactaacc cctctgcaaa ttcttcaggg ccttgaacag caccctggaa gggttcatag
     6001 aataaagctt caactccttc agacagacct ctaattaatc caaatgggtt tccaagtaca
     6061 tctaacccca atacaaggac atacatctgt ttcaagaact gttcactgta atgcctaaca
     6121 acactccata taagctgatc tctcttgtag aactgatatc gaatttcata ataagcaagt
     6181 ttgaatataa ggtcatccac atcagtcaga gtagcaccta tgcttttcaa cagcaagttg
     6241 acagaatgaa ctgcaaacat ttcctgtttt tctttgtctg attcttcacc tccggaaccc
     6301 aaagacaaac tcaaatgcaa cttcacagga gaaatatgga aatgttcaaa gaaactaaga
     6361 attgacatat cagtcattga agtctccatt aattctgcat ttagagcatc aatatcttgt
     6421 tggattaact ttgtccgtct tctttcagct tcagggtctg ttgttggggt aaacagtgca
     6481 ataatagctc ctagaaaccc ttgatcaatt tttaaggcca tttcctgaat gaggaccata
     6541 aaatacttga actgtaagac tttactgtac tcattaaatc ttgtgatgac actcacatca
     6601 atgaaaggct tgggctctga atctaaagca atagattttg gaggggcaac aggatgaaat
     6661 acaacaggga acattgcacc tggtaactga ttatcaacct gaagccagta caacctggcc
     6721 cttaaacttc tctggtgaga agactgctta aattcaatct gaattcctga taaaaagtct
     6781 cgtttaatag ggctacgaat agggaggcgc atttccattg gatctttatc aaaattgacc
     6841 tcaaaattat tatctagctt aatccagcca tggtctcttg atatttgatg tttctgatag
     6901 gattgttcca ataagattat ctgcttttga ctaaatggct tccatttctg ctttggtttc
     6961 acctcccaaa caacaccaga actggttatc ccaatatagg aaacttcctg cttgctttca
     7021 ttgttaacca gtgaaagccc aagactgtgg agagacaagg ttatttcata atcagcctgt
     7081 tccatttctt ctgcctgcag tgctttggaa accaaggcaa catcatcggt gaaaagcaaa
     7141 actctctggc gcccatccag aaatgatacc cagtgtatct ggatgtttgc atcatatgga
     7201 aactgtccac atccatcctt taacagatca tgttccccaa catttgctgc atatgtccat
     7261 gtaagttttc tggtaccagt aggatctgcc caggcaaaaa gtcgagcctg tcttggcagc
     7321 aagaccattt cttctggtga cccactctgt ttgtatgtga ggatgtccca tggtgtatgg
     7381 ttcattatca aggcaggtgc agatccctca tggtaatcag aaaaagttat gacagttgaa
     7441 tgttcggcag tgtttacatc caccaagata cccccattca gatcttctaa gctcaataaa
     7501 gtgccattat cctgtcggtt ataaaagaat ggtttggaag atccttcaca gcccaccact
     7561 ctcacacaaa gtttgcctga caaactttct ggccaaaatg gaaggcactc tgaagaagca
     7621 atatagttcc atttattagt tggcattgag ccatcagatg caatctcgcc aacttctagt
     7681 tctaatgatg acttgtttgc aatggtacaa aagggagtca gggtaactat tcgtgaaagg
     7741 ttgaaactgc tcattttgat gctaacacca accaggtact ccatattgtt ggcaggacac
     7801 ttcacacacc cataacttcc cactgtatcc aatgagaaac tactggacca ggcactggtt
     7861 gaaattttta attgtacctt atttttagta aaaatgttct tcttcttgaa agaaaataaa
     7921 ataatatccc tgaaatcagc tggatgtttc acatgaatat cttctgaacg atactggaga
     7981 acccgggtag tcttgttgat taaccaatag ggactaaaga cagacagcac catccggctg
     8041 ccaattctcc tgacgtggac tgacaggtcg actgtcgtca cttctgtgga gtcagaagaa
     8101 aaacacacag gaaagaattc tggtagtgta tcacgtatgc ggaaatgtcc attccagttt
     8161 ttgccctggt atttcaccag gactaattcc attatttcac cactgattct cgaatgcaga
     8221 acatcagcag tactgccttc tgccagctca tgagtttctg ctgttccctc aagtaaatat
     8281 cttagggaat atgggagaag attccgcaaa gtgagagaag gataaagatg aataatgtaa
     8341 gctacatccc agtcttcccc atgtgtacat atgtagctca attcatcagg cagagcaact
     8401 gtattcacta tgagaggtaa gaagctgact tctactgatg gacactgcaa catgcatctg
     8461 acttccctgc tcctatgaag ttcttccttc caggaaatat aagtggtaga ttctttgtac
     8521 tgatgctcta agattccagc tggctggata aacaattgac atctatatga atctaaagga
     8581 acatggaact cctcttcagg tctggctatc ccaatgcgct ccaatagctt aacattctta
     8641 acaaatttat agatgataaa tgcaatggag aaatggtttt tgatctgtag aggagagcga
     8701 agggtaatt
//
