LOCUS       sequence-444787-        6623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synchronize the trafficking of CD9 from the ER..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6623)
  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 6623)
  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_CD9-SBP-mCherry
FEATURES             Location/Qualifiers
     source          1..6623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..83
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(320..339)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          322..551
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1315..1334)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1316..1334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1379..1582)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1408..1428)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1583..1962
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2134..2153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2228..2332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2333..2401,2402..3193)
                     /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     complement(2551..2570)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3364..3952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3853..3872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4106..4123
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4240..4261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4276..4281,4282..4299,4300..4306)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4314..4330
                     /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)."
     primer_bind     4319..4341
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4338..4354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4338..4354
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4456..4577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4521..4540
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5326..6030)
                     /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(5601..5620)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5870..5888
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(6040..6150)
                     /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"
     misc_feature    join(6287..6295,6296..6298,6299..6623,1..250)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6460..6479)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 agacgttgtt tgtcttcaag aagcttccag aggaactgct tccttcacga cattcaacag
       61 accttgcatt cctttggcga gaggggaaag acccctagga atgctcgtca agaagacagg
      121 gccaggtttc cgggccctca cattgccaaa agacggcaat atggtggaaa ataacatata
      181 gacaaacgca caccggcctt attccaagcg gcttcggcca gtaacgttag ggggggggga
      241 gggagagggg gggggggaga ggggcggaat tggccgccct agatgcatgc tcgacctgca
      301 gttggacctg ggagtggaca cctgtggaga gaaaggcaaa gtggatgtca ttgtcactca
      361 agtgtatggc cagatctcaa gcctgccaca cctcaagctt gacaacaaaa agattgtctt
      421 ttctgaccag atggacgcgg ccaccctcaa aggcatcacc gcgggccagg tgaatatcaa
      481 atcctcctcg tttttggaaa ctgacaatct tagcgcagaa gtcatgcccg cttttgagag
      541 ggagtactca ccccaacagc tggccctcgc agacagcgaa ttaattccag cacactggat
      601 cagttatcta tgcggccgct cacagttcat ctttcggtcc gagctgctgg acggcatcca
      661 gaggattgcc gttgttgact ccggcttttt tggcggcatc gattgaagcg gcggagggtt
      721 tcactttggt gaatgtgtca tgtcccacca gggtggattt ccaagcattt gcctcggttg
      781 tgccagatgt gagcagccac tgtgtattga ttctagcctc agcgcctccc acgtactgtc
      841 cactccatgt tgtggcagaa tgagcattcc ggtagttgtt tttccaagcg acagtccatc
      901 ccagggcagt cccagatccg tcggtggcag gggcactatc gtatcggcca gtgaggacgt
      961 accgtgattc ggcgtttccc acagcagatt cgtatgttcc ggtgagggct ccatcggctc
     1021 cagcagtcac aataaaggtg ctcccgagct gattgtacca tgttccagta atgccagcct
     1081 cggcagcgga cacctgagct tttgagtctt tgctagggtc gtttaaaccg gcgcccgagc
     1141 tggttcctgt cgccttctca ctgtgactat ggctgaggct ctggccctgg tgcaggagga
     1201 gtccccagag gagccacagg gcaagacgga cgcatacatc cataccggtt aacaggcctt
     1261 aagcgctagc taggccgcag atatcgatcc gagctcggta ccaagcttgg gtctccctat
     1321 agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag
     1381 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     1441 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     1501 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     1561 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     1621 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     1681 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     1741 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     1801 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     1861 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     1921 ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc
     1981 gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg
     2041 tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca
     2101 cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag
     2161 tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg
     2221 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2281 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2341 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2401 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2461 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2521 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2581 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2641 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2701 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2761 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2821 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2881 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2941 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3001 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3061 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3121 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3181 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3241 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3301 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3361 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3421 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3481 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3541 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3601 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3661 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3721 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3781 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3841 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3901 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3961 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4021 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4081 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     4141 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     4201 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     4261 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     4321 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga
     4381 ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     4441 atgcactcga cgcgttaaga tacattgatg agtttggaca aaccacaact agaatgcagt
     4501 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     4561 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     4621 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     4681 atcagttatc tagagttaat taataattgg ccctcggccc tagaccatct cgcggttcct
     4741 gcggatagca cagcacaaga tcatactgaa gatcatgcca aatatcatga ccacggcaat
     4801 gccgatgccc actgcgccga tgatgtggaa tttattgtcg aagacctctt tgatggcatc
     4861 aggacaggac ttcacggtga aggtttcgag tacgtccttc ttggggcaga tgtctgagat
     4921 aaactgttcc acgcccccag ccaaaccaca gcagttcaac gcatagtgga tggctttcag
     4981 cgtttcccgc tggggctcat ccttggtttt cagcttgttg taggtgtcct tgtaaaactc
     5041 ctggacttcc ttaatcacct catccttgtg ggaatatccc cagatggccg cagctatttc
     5101 aatggcgaat atcaccaaga ggaagccgaa gaacagtccc agcatgcact gggactcctg
     5161 cacagccccg cagcagccca ggaagcccac cagcatcatg agggcgccgg ctccgatcag
     5221 aatatagact cctgtgtaga agctggaatt attattatta gtttcttgct cgaagatgct
     5281 cttggtctga gaagatctga ttcgagctcc accggttggc cggcccttgt acagctcgtc
     5341 catgccgccg gtggagtggc ggccctcggc gcgttcgtac tgttccacga tggtgtagtc
     5401 ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg taggcgccgg gcagctgcac
     5461 gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc cgtccttcag
     5521 cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca tccgctcgga
     5581 ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggagg ggaagttggt
     5641 gccgcgcagc ttcaccttgt agatgaactc gccgtcctgt agggaggagt cctgggtcac
     5701 ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa
     5761 ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct tggagccgta
     5821 catgaactga ggggacagga tgtcccaggc gaagggcagg gggccaccct tggtcacctt
     5881 cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa
     5941 ctcgtggccg ttcacggagc cctccatgtg caccttgaag cgcatgaact ccttgatgat
     6001 ggccatgtta tcctcctcgc ccttgctcac acctgcaggt ggttcacgtt gaccttgtgg
     6061 gtgatgctcc agtcgtgcac gaagttgttc aagttcccca gccagtcctt caacaacgtg
     6121 tccacctcgc caaccagtgg tcttctcgtc gaattcgtcg aatcggagcc atagtccaat
     6181 ggcaaggaca gcaatcccgg caagccagaa gatgaagtta aatccgaaca gcaggtattt
     6241 gatgcacttg gtgcctcctt tgaccggagg catggcgcgc ctcccgggtt gtggcaagct
     6301 tatcatcgtg tttttcaaag gaaaaccacg tccccgtggt tcggggggcc tagacgtttt
     6361 tttaacctcg actaaacaca tgtaaagcat gtgcaccgag gccccagatc agatcccata
     6421 caatggggta ccttctgggc atccttcagc cccttgttga atacgcttga ggagagccat
     6481 ttgactcttt ccacaactat ccaactcaca acgtggcact ggggttgtgc cgcctttgca
     6541 ggtgtatctt atacacgtgg cttttggccg cagaggcacc tgtcgccagg tggggggttc
     6601 cgctgcctgc aaagggtcgc tac
//
