LOCUS       sequence-500760-        5224 bp    DNA     circular SYN 23-FEB-2026
DEFINITION  This plasmid can be used to quantify CD9 by luminescence signal upon
            providing substrate, in particular in the secretome, especially in 
            the extracellular vesicles of cells expressing this construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5224)
  AUTHORS   Grisard E, Nevo N, Lescure A, Doll S, Corbe M, Jouve M, Lavieu G, 
            Joliot A, Nery ED, Martin-Jaular L, Thery C
  TITLE     Homosalate boosts the release of tumour-derived extracellular 
            vesicles with protection against anchorage-loss property.
  JOURNAL   J Extracell Vesicles. 2022 Jul;11(7):e12242. doi: 
            10.1002/jev2.12242.
  PUBMED    35790086
REFERENCE   2  (bases 1 to 5224)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 23, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Extracell
            Vesicles."; date: "2022-07"; volume: "11(7):e12242. doi"; pages: " 
            10.1002/jev2.12242"
COMMENT     Sequence Label: Nluc CD9
FEATURES             Location/Qualifiers
     source          1..5224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             109..903
                     /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             109..903
                     /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(163..182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     773..792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1091..1110)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1135..1182
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1511..2099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2000..2019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2274..2577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2578..2781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2732..2752
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2819..3331
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    4225..4346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4262..4281)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4316..4335
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4353..4808)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4490..4511)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4702..4721
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4835..4939
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4941..5224,1..74)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4941..5132
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4964..4984)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5149..5224,1..60)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
       61 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
      301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
      661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
      841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
      901 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
      961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     1081 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     1141 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     1201 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     1261 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     1321 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     1381 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1441 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1621 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1741 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1801 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1861 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1921 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1981 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2041 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2101 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2161 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     2221 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2281 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2341 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2401 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2461 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2521 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2581 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2641 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2701 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2761 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtat
     2821 ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga
     2881 ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac
     2941 tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat
     3001 catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     3061 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     3121 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     3181 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     3241 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     3301 cggctggcgg ctgtgcgaac gcattctggc gctcgagctc aagcttccac cggtcaaagg
     3361 aggcaccaag tgcatcaaat acctgctgtt cggatttaac ttcatcttct ggcttgccgg
     3421 gattgctgtc cttgccattg gactatggct ccgattcgac tctcagacca agagcatctt
     3481 cgagcaagaa actaataata ataattccag cttctacaca ggagtctata ttctgatcgg
     3541 agccggcgcc ctcatgatgc tggtgggctt cctgggctgc tgcggggctg tgcaggagtc
     3601 ccagtgcatg ctgggactgt tcttcggctt cctcttggtg atattcgcca ttgaaatagc
     3661 tgcggccatc tggggatatt cccacaagga tgaggtgatt aaggaagtcc aggagtttta
     3721 caaggacacc tacaacaagc tgaaaaccaa ggatgagccc cagcgggaaa cgctgaaagc
     3781 catccactat gcgttgaact gctgtggttt ggctgggggc gtggaacagt ttatctcaga
     3841 catctgcccc aagaaggacg tactcgaaac cttcaccgtg aagtcctgtc ctgatgccat
     3901 caaagaggtc ttcgacaata aattccacat catcggcgca gtgggcatcg gcattgccgt
     3961 ggtcatgata tttggcatga tcttcagtat gatcttgtgc tgtgctatcc gcaggaaccg
     4021 cgagatggtc tagactagtg gatccgagct cggtaccaag cttcgaattc tgcagtcgac
     4081 ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag ccataccaca
     4141 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     4201 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4261 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4321 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     4381 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4441 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4501 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4561 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4621 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4681 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4741 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4801 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4861 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4921 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4981 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5041 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5101 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5161 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5221 gagg
//
