LOCUS       sequence-367335-        6287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human MOSPD2 (MSP domain from residue 282 to 490; 
            mutant R404D/L406D, unable to bind FFAT motifs), fused to a 6HIS 
            tag, in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6287)
  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 6287)
  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: pET15b MOSPD2 RD/LD [282-490]
FEATURES             Location/Qualifiers
     source          1..6287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      583..1171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1072..1091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1325..1342
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1357..1496
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1422..1441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1582..1604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1598..1789)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4144..4163
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4183..4260)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4466..4485
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4569..4588
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4569..4587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4588..4612
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             4627..4649
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5286..5303
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5357..5375)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5371..5418
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5593..5598,5599..5615,5616..5621))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5648..5666)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5734..5838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5839..5907,5908..6287,1..412)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6057..6076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
       61 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
      121 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
      181 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
      241 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
      301 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
      361 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
      421 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
      481 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
      541 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
      601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
      661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
      721 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      901 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      961 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1021 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1081 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1141 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1201 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1261 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     1321 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     1381 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     1441 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     1501 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     1561 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     1621 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     1681 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     1741 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     1801 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     1861 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     1921 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     1981 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2041 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2101 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2161 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2221 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2281 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     2341 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     2401 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     2461 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     2521 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt
     2581 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     2641 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     2701 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     2761 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     2821 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     2881 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     2941 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3001 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     3061 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3121 gggaaactgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3181 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     3241 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     3301 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     3361 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     3421 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     3481 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     3541 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     3601 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     3661 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     3721 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     3781 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     3841 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     3901 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     3961 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     4021 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     4081 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     4141 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     4201 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     4261 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     4321 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     4381 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     4441 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     4501 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     4561 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     4621 aataattttg tttaacttta agaaggagat ataccatgat ttctaatgaa gaacaaacac
     4681 ctcttcttaa aaagattaac ccaaccgaat ctacttccaa agcagaagaa aatgaaaaag
     4741 ttgattcaaa agtgaaagct ttcaagaaac cattgagtgt atttaaaggc cccttactac
     4801 acatcagccc agcagaagaa ctgtactttg gaagtacaga atccggagag aagaaaacct
     4861 taatagtgtt gacaaatgta actaaaaata tagtggcatt taaggtgaga acaacagctc
     4921 cagaaaaata cagagtcaag ccaagcaata gcagctgtga cccgggtgca tcagtggata
     4981 tagttgtgtc tccccatggg ggtttaacag tctctgccca agacgatttt gacataatgg
     5041 ctgcagaaat ggaacagtca tctggcacag gcccagcaga attaactcag ttttggaaag
     5101 aagttcccag aaacaaagtg atggaacata ggttaagatg ccatactgtt gaaagcagta
     5161 aaccaaacac tcttacgtta aaagacaatg ctttcaatat gtcagataaa accagtgaag
     5221 atatatgtct acaactcagt cgtttactag aaagcaatag gaagcttgaa gaccaagttc
     5281 agcgtcatca tcatcatcat cactgaagat ccggctgcta acaaagcccg aaaggaagct
     5341 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     5401 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg
     5461 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     5521 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     5581 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga
     5641 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5701 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5761 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5821 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5881 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5941 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6001 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6061 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     6121 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6181 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6241 ttacttctga caacgatcgg aggaccgaag gagctaaccg ctttttt
//
