LOCUS       sequence-472716-        6638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the VAB domain of VPS13C in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6638)
  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 6638)
  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-VAB(VPS13C)
FEATURES             Location/Qualifiers
     source          1..6638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3426..3547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3463..3482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3517..3536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3554..4009)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3691..3712)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3903..3922
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4036..4140
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4142..4499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4142..4333
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4165..4185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4350..4485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4534..5328
                     /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             4534..5328
                     /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(4588..4607)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5198..5217
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5516..5535)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5560..5607
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5936..6524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6425..6444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 atgagagccc ccttcaccgt gaagaatgcc gtgggcgtgc caatcaaggt gaagcccaac
     1381 tgtaatctgc gcgtgatggg ctttcctgag aagagcgaca tcttcgacgt ggatgccggc
     1441 cagaacctgg agctggagta cgcctccatg gtgccatcta gccagggcaa tctgtccatc
     1501 ctgtctaggc aggagtcctc tttctttacc ctgacaatcg tgccccacgg ctataccgag
     1561 gtggccaaca tccctgtggc cagaccaggc cggagactgt acaatgtgag gaaccccaat
     1621 gccagccact ccgactctgt gctggtgcag atcgatgcca ccgagggcaa caaagtgatc
     1681 accctgcgca gccctctgca gatcaagaat cacttttcca tcgccttcat catctataag
     1741 ttcgtgaaga acgtgaagct gctggagaga atcggcatcg ccaggcctga ggaggagttt
     1801 cacgtgccac tggacagcta caggtgccag ctgttcatcc agccagccgg catcctggag
     1861 caccagtata aggagtccac cacatacatc tcttggaagg aggagctgca cagaagcagg
     1921 gaggtgcgct gcatgctgca gtgtccatct gtggaggtga gctttctgcc cctgatcgtg
     1981 aacaccgtgg ccctgcccga tgagctgtcc tacatctgta cacacggcga ggactgggat
     2041 gtggcctata tcatccacct gtaccctagc ctgaccctgc gcaatctgct gccatattcc
     2101 ctgaggtacc tgctggaggg aacagcagag acacacgagc tggcagaggg aagcacagca
     2161 gacgtgctgc acagccggat ctccggagag atcatggagc tggtgctggt gaagtaccag
     2221 ggcaagaact ggaatggcca ctttcgcatc cgggataccc tgcccgagtt ctttcccgtg
     2281 tgcttcagct ccgactccac agaggtgacc acagtggatc tgtctgtgca cgtgaggagg
     2341 atcggcagcc ggatggtgct gagcgtgttc agcccttatt ggctgatcaa caagaccaca
     2401 cgcgtgctgc agtaccggtc cgaggacatc cacgtgaagc acccagccga ctttagagat
     2461 atcatcctgt tcagcttcaa gaagaagaac atcttcacaa agaataaggt gcagctgaag
     2521 atcagcacct ccgcctggtc tagctccttt tccctggata ccgtgggctc ttatggctgc
     2581 gtgaagtgtc ccgccaacaa tatggagtac ctggtgggcg tgtctatcaa gatgtctagc
     2641 tttaacctga gcaggatcgt gaccctgaca ccattctgca ccatcgccaa taagtcctct
     2701 ctggagctgg aggtcggaga gatcgcatcc gacggatcta tgcccacaaa caagtggaat
     2761 tatatcgcca gctccgagtg tctgcctttc tggccagagt ctctgagcgg caagctgtgc
     2821 gtgcgggtgg tgggatgtga gggctctagc aagcccttct tttacaacag acaggataat
     2881 ggcaccctgc tgagcctgga ggacctgaac ggcggcatcc tggtggatgt gaatacagcc
     2941 gagcacagca cagtgatcac cttttccgac tatcacgagg gctctgcccc tgccctgatc
     3001 atgaaccaca ccccatggga tatcctgaca tacaagcagt ccggctctcc cgaggagatg
     3061 gtgctgctgc ctaggcaggc aaggctgttt gcatgggcag accctaccgg aacaaggaag
     3121 ctgacctgga catatgcagc aaacgtggga gagcacgacc tgctgaagga tggctgtggc
     3181 cagttcccat acgacgccaa tatccagatc cactgggtga gctttctgga tggcagacag
     3241 agggtgctgc tgttcaccga ctgatgaatt ctgcagtcga cggtaccgcg ggcccgggat
     3301 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     3361 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3421 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3541 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3601 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3661 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3721 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3781 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3841 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3901 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3961 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4021 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4081 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4141 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4201 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4261 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4321 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4381 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4441 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4501 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4561 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4621 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4681 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4741 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4801 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4861 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4921 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4981 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5041 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5101 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5161 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5221 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5281 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5341 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5401 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5461 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5521 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5581 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5641 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5701 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5761 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5821 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5881 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5941 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6001 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6061 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6121 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6181 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6241 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6301 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6361 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6421 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6481 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6541 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6601 cccctgattc tgtggataac cgtattaccg ccatgcat
//
