LOCUS       sequence-447859-        5544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human VAP-B (mutant K87D/M89D, unable to bind FFAT 
            motifs) fused to mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5544)
  AUTHORS   Di Mattia T, Wilhelm LP, Ikhlef S, Wendling C, Spehner D, Nomine Y, 
            Giordano F, Mathelin C, Drin G, Tomasetto C, Alpy F
  TITLE     Identification of MOSPD2, a novel scaffold for endoplasmic reticulum
            membrane contact sites.
  JOURNAL   EMBO Rep. 2018 Jul;19(7). pii: embr.201745453. doi: 
            10.15252/embr.201745453. Epub 2018 Jun 1.
  PUBMED    29858488
REFERENCE   2  (bases 1 to 5544)
  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"; journalName: "EMBO Rep.";
            date: "2018-07"; volume: "19(7). pii: embr.201745453. doi"; pages: "
            10.15252/embr.201745453. Epub 2018 Jun 1"
COMMENT     Sequence Label: pmCherryC1 VAP-B KD/MD
FEATURES             Location/Qualifiers
     source          1..5544
                     /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"
     CDS             591..620
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     CDS             join(624..632,633..647)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             660..1367
                     /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(805..823)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1073..1092
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1407..1433
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2332..2453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2369..2388)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2423..2442
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2460..2915)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2597..2618)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2809..2828
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2942..3046
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3048..3405
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3048..3239
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3071..3091)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3256..3391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3440..4234
                     /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             3440..4234
                     /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(3494..3513)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4104..4123
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4422..4441)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4466..4513
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4842..5430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5331..5350
                     /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 gctagcatga
      601 ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat cccgccacca
      661 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      721 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      781 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      841 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      901 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      961 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     1021 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     1081 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     1141 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     1201 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     1261 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1321 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtcc ggactcagat
     1381 ctcgagctca agcttcgaat tgtgcctacc catatgacgt cccggactac gccggatcca
     1441 tggcgaaggt ggagcaggtc ctgagcctcg agccgcagca cgagctcaaa ttccgaggtc
     1501 ccttcaccga tgttgtcacc accaacctaa agcttggcaa cccgacagac cgaaatgtgt
     1561 gttttaaggt gaagactaca gcaccacgta ggtactgtgt gaggcccaac agcggaatca
     1621 tcgatgcagg ggcctcaatt aatgtatctg tgatgttaca gcctttcgat tatgatccca
     1681 atgagaaaag taaacacgac tttgacgtcc agtctatgtt tgctccaact gacacttcag
     1741 atatggaagc agtatggaag gaggcaaaac cggaagacct tatggattca aaacttagat
     1801 gtgtgtttga attgccagca gagaatgata aaccacatga tgtagaaata aataaaatta
     1861 tatccacaac tgcatcaaag acagaaacac caatagtgtc taagtctctg agttcttctt
     1921 tggatgacac cgaagttaag aaggttatgg aagaatgtaa gaggctgcaa ggtgaagttc
     1981 agaggctacg ggaggagaac aagcagttca aggaagaaga tggactgcgg atgaggaaga
     2041 cagtgcagag caacagcccc atttcagcat tagccccaac tgggaaggaa gaaggcctta
     2101 gcacccggct cttggctctg gtggttttgt tctttatcgt tggtgtaatt attgggaaga
     2161 ttgccttgta gcaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga
     2221 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2281 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2341 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2401 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     2461 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2941 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3001 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3061 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3121 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3421 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3481 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3541 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3601 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3661 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3721 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3781 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3841 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3901 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3961 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4021 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4081 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4141 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4201 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4261 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4321 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4381 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4441 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4501 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4561 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4621 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4681 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5521 gataaccgta ttaccgccat gcat
//
