LOCUS       sequence-442318-        6649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synchronize the trafficking of CD63 from the ER..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6649)
  AUTHORS   Mathieu M, Nevo N, Jouve M, Valenzuela JI, Maurin M, Verweij FJ, 
            Palmulli R, Lankar D, Dingli F, Loew D, Rubinstein E, Boncompain G, 
            Perez F, Thery C
  TITLE     Specificities of exosome versus small ectosome secretion revealed by
            live intracellular tracking of CD63 and CD9.
  JOURNAL   Nat Commun. 2021 Jul 19;12(1):4389. doi: 10.1038/s41467-021-24384-2.
  PUBMED    34282141
REFERENCE   2  (bases 1 to 6649)
  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: "Nat 
            Commun."; date: "2021-07-19"; volume: "12(1):4389. doi"; pages: " 
            10.1038/s41467-021-24384-2"
COMMENT     Sequence Label: Str-KDEL_CD63-SBP-mCherry
FEATURES             Location/Qualifiers
     source          1..6649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     867..886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        867..885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1650..1879
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1862..1881
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1951..2525,2526..2528,2529..2537)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2118..2135)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2345..2364
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2680..2790
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             2800..3504
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2942..2960)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3210..3229
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4273..4394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4310..4329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4496..4512)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4496..4512)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4509..4531)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4520..4536
                     /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)."
     promoter        complement(join(4544..4550,4551..4568,4569..4574))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4589..4610
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4727..4744)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4898..5486)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4978..4997)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5657..6448,6449..6517))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6280..6299
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6518..6622)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
      901 taccgagctc ggatcgatat ctgcggccta gctagcgctt aaggcctgtt aaccggtatg
      961 gatgtatgcg tccgtcttgc cctgtggctc ctctggggac tcctcctgca ccagggccag
     1021 agcctcagcc atagtcacag tgagaaggcg acaggaacca gctcgggcgc cggtttaaac
     1081 gaccctagca aagactcaaa agctcaggtg tccgctgccg aggctggcat tactggaaca
     1141 tggtacaatc agctcgggag cacctttatt gtgactgctg gagccgatgg agccctcacc
     1201 ggaacatacg aatctgctgt gggaaacgcc gaatcacggt acgtcctcac tggccgatac
     1261 gatagtgccc ctgccaccga cggatctggg actgccctgg gatggactgt cgcttggaaa
     1321 aacaactacc ggaatgctca ttctgccaca acatggagtg gacagtacgt gggaggcgct
     1381 gaggctagaa tcaatacaca gtggctgctc acatctggca caaccgaggc aaatgcttgg
     1441 aaatccaccc tggtgggaca tgacacattc accaaagtga aaccctccgc cgcttcaatc
     1501 gatgccgcca aaaaagccgg agtcaacaac ggcaatcctc tggatgccgt ccagcagctc
     1561 ggaccgaaag atgaactgtg agcggccgca tagataactg atccagtgtg ctggaattaa
     1621 ttcgctgtct gcgagggcca gctgttgggg tgagtactcc ctctcaaaag cgggcatgac
     1681 ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc
     1741 ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc
     1801 aagcttgagg tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac
     1861 tttgcctttc tctccacagg tgtccactcc caggtccaac tgcaggtcga gcatgcatct
     1921 agggcggcca attccgcccc tctccccccc cccctctccc tccccccccc ctaacgttac
     1981 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     2041 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     2101 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     2161 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     2221 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     2281 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     2341 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     2401 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     2461 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     2521 ataagcttgc cacaacccgg gaggcgcgcc atggcggtgg aaggaggaat gaaatgtgtg
     2581 aagttcttgc tctacgtcct cctgctggcc ttttgcgcct gtgcagtggg actgattgcc
     2641 gtgggtgtcg gggcacagag atctctagcc accgaattcg acgagaagac cactggttgg
     2701 cgaggtggac acgttgttga aggactggct ggggaacttg aacaacttcg tgcacgactg
     2761 gagcatcacc cacaaggtca acgtgaacca cctgcaggtg tgagcaaggg cgaggaggat
     2821 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     2881 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2941 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3001 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3061 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3121 gtggtgaccg tgacccagga ctcctcccta caggacggcg agttcatcta caaggtgaag
     3181 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     3241 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3301 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3361 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3421 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3481 ggcggcatgg acgagctgta caagggccgg ccaaccggtg gagctcgaat cagatctctt
     3541 gtcctgagtc agaccataat ccagggggct acccctggct ctctgttgcc agtggtcatc
     3601 atcgcagtgg gtgtcttcct cttcctggtg gcttttgtgg gctgctgcgg ggcctgcaag
     3661 gagaactatt gtcttatgat cacgtttgcc atctttctgt ctcttatcat gttggtggag
     3721 gtggccgcag ccattgctgg ctatgtgttt agagataagg tgatgtcaga gtttaataac
     3781 aacttccggc agcagatgga gaattacccg aaaaacaacc acactgcttc gatcctggac
     3841 aggatgcagg cagattttaa gtgctgtggg gctgctaact acacagattg ggagaaaatc
     3901 ccttccatgt cgaagaaccg agtccccgac tcctgctgca ttaatgttac tgtgggctgt
     3961 gggattaatt tcaacgagaa ggcgatccat aaggagggct gtgtggagaa gattgggggc
     4021 tggctgagga aaaatgtgct ggtggtagct gcagcagccc ttggaattgc ttttgtcgag
     4081 gttttgggaa ttgtctttgc ctgctgcctc gtgaagagta tcagaagtgg ctacgaggtg
     4141 atgtagttaa ttaactctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     4201 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     4261 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4321 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4381 atcaatgtat cttaacgcgt cgagtgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4441 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4501 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4561 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4621 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4681 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4741 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4801 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4861 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4921 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4981 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5041 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5101 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5161 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5221 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5281 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5341 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5401 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5461 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5521 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5581 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5641 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5701 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5761 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5821 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5881 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5941 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6001 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6061 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6121 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6181 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6241 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6301 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6361 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6421 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6481 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6541 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6601 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctga
//
