LOCUS       sequence-472704-        7106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses OSBP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7106)
  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 7106)
  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-OSBP
FEATURES             Location/Qualifiers
     source          1..7106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(135..929)
                     /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(135..929)
                     /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(246..265)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     856..875
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(964..1321)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      978..1113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1130..1321
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1278..1298
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1323..1427)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1454..1909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1541..1560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1751..1772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1916..2037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1927..1946)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1981..2000
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      4583..4592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4612..5319)
                     /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(4887..4906)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5156..5174
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5316..5325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5363..5566)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5392..5412)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5567..5870
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6045..6633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6125..6144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6962..7009
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     7034..7053
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
       61 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
      121 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
      181 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
      241 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
      301 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
      361 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac
      421 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
      481 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
      541 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
      601 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
      661 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
      721 agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc
      781 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
      841 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
      901 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg
      961 atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     1021 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     1081 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     1141 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     1201 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     1261 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     1321 ggactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1381 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1441 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1501 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1561 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1621 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1681 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1741 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1801 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1861 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgccttaaga
     1921 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1981 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2041 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2101 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagttatc tagatccggt
     2161 ggatcctcag aaaatgtccg ggcatgagct ccagtcctgt ttttctttac actcccagta
     2221 ttctccccta taaatatggg ttaactcctt ggtaacaggg tccttcttcc gctcaaacca
     2281 cagtgcctta tagggatcat atggtgtgcc atcctctgtg gctttcatag cttccgcttc
     2341 tctcttcttt ctggaaagtc tttgtttttc ctccaggcgc tgcttctccg catttgcttc
     2401 atcccagcgt ccattttcca tcagtctctg gtcaggtcgt aaccggctgt ctgtgggggc
     2461 agtgccactt tcccaagcat tgagagtcag agcaagctct gagaagtagt acatgttttc
     2521 tgcattcttc ggtaaaggat tccttttcca cagcatgacc ctgctttcct ctgcttcatg
     2581 gcctctctgt cgagcatcac ccccattttc cccaatgact ggctgtactt tgaaacattc
     2641 cattttctca tcccacgtcc ccagaagagc aaagtggact tttcctgatg gatctgtcac
     2701 ttcccccgtc acctttcttg ctacatcccg agagaagtag ctataaggaa caaatttaag
     2761 attacacttg tctcctgtct tgtgattcac aatatcaatt tcgccagact gatctatcca
     2821 caacttgccc acaataatgt tgtgtacagt tgtggtaact ttcttccaag tgtagtggtg
     2881 cccagttgca tggaaaatac aatgaatggt accgaggggc ataatggaga ggtatttgcc
     2941 tcgaaacttg ctggtgattt tgatttcctg acgcaatgtc cagccatttt tggactcagc
     3001 atggtgcgca gcagcagggg gatgatgact cacctgttca cagagggatc ggtacccatt
     3061 ctcctctaat cggtccagct caaaggtctc cccaagcagt gggttgaatg gcttactggt
     3121 gcggaagaca gtagtggagt aggaggacac ggtgaaagct gcaacataac agagctgttc
     3181 tagagaattc tcacattttg cagctcggtc taacagctca tggtattcca gatcttcagt
     3241 aaggcgctga agcatggaca agggctcatt aaagtttacc ggcatgggga tcttagagag
     3301 ttcttttcca atgcagttct tcatgatgct ccataaattg aggctatagt ttggcttgta
     3361 tggtattctg gttctctttt cctttttggt ctcctccagc tgatgcttgt actgttcatc
     3421 aaggctgatg tcactgctgg ctccactgat attgctgcca gtacgtttgt ggcccaaatt
     3481 ttcaggcatg gtgatgatct caggtgcatc aaaaaattca ttctcatcat cttcatcgct
     3541 catgtcccct ttgccagagc agcactgatc tttaccagaa cccacattgc caggagtgtt
     3601 tgccggcagc accgtggctc ctcggaaggc cctctccagg tgattatgct gcttcgccag
     3661 ctgctcgagg gtttcttcca gtcggatacg ctggtctctt tcatactgta gtgacttttg
     3721 ccatttttta ctatgggtct gggctaacat gaggaaatct ctgcaggcgt tgatcatggc
     3781 attggatgtt atcctaaaga gtgtggctcg ttcgttgacc tgtttgatct tttcattgct
     3841 ctcagcaggc aacttcaggg actccagctc actgagagaa cgctgcagag ctgtgccatg
     3901 cttagctatc aagtcattgc acgtgctcaa gtcctctact ttgctagaga gggtccgaag
     3961 ggtattctgc agctcagtct tgtcagtttg tgagacagac tcttcatctc ctgattcatc
     4021 tgactctgcc agcatcttca cagctttggc cttggccagt tccagggccg tcacccagcg
     4081 ctgccgctca acttctgaac tagctttcag atggtaggtc tgagcacccc cattggaaat
     4141 gatgaagttg caggagtcct ccacggtgat gttggctgtg gcgaggttga tggtaccacg
     4201 gcaggtatgt ctcatttctg cctttgatct gtagtagctc aggagcccgt tgctcagcac
     4261 gaaccatcgc cgctggtagc ctttgatata attggtccat ttgaagagcc agccctctcg
     4321 agccgagccc gaacccccag cgcccgagcc gcccgagccc ccagtcggcg gcgcaggggc
     4381 cgggccagcc gccgccactc ccccggcccc cgggcccggg cctcccgccg ccgccgcgac
     4441 caccgtccct gacgcggccc cggagcctgg ccccgcatct ccgcggccgc cgcctcctcc
     4501 caccactggg ggaccggcgc cgccgccgcc aagtgctgca atggctgccg ggcctggccc
     4561 caccactcct ctcagctccg tcgccgccat ggaagcttga gctcgagatc tcttgtacag
     4621 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
     4681 gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag
     4741 ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc
     4801 cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg
     4861 ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa
     4921 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     4981 ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc
     5041 ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     5101 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     5161 caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat
     5221 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     5281 gatgatggcc atgttatcct cctcgccctt gctcaccatg gtggcaccgg tagcgctagc
     5341 ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
     5401 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     5461 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     5521 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     5581 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     5641 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     5701 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     5761 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     5821 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     5881 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg
     5941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6781 tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg
     6841 ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc
     6901 caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta
     6961 tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
     7021 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag
     7081 aactccagca tgagatcccc gcgctg
//
