LOCUS       sequence-359687-        5278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing COM-CD63-COM fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5278)
  AUTHORS   Yao X, Lyu P, Yoo K, Yadav MK, Singh R, Atala A, Lu B
  TITLE     Engineered extracellular vesicles as versatile ribonucleoprotein 
            delivery vehicles for efficient and safe CRISPR genome editing.
  JOURNAL   J Extracell Vesicles. 2021 Mar;10(5):e12076. doi: 
            10.1002/jev2.12076. Epub 2021 Mar 16.
  PUBMED    33747370
REFERENCE   2  (bases 1 to 5278)
  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: "J Extracell
            Vesicles."; date: "2021-03"; volume: "10(5):e12076. doi"; pages: " 
            10.1002/jev2.12076. Epub 2021 Mar 16"
COMMENT     Sequence Label: pCOM-CD63-COM
FEATURES             Location/Qualifiers
     source          1..5278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..228
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     179..199
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             498..1211
                     /codon_start=1
                     /product="human CD63 tetraspanin"
                     /label=CD63
                     /note="enriched in intraluminal vesicles of late endosomes 
                     (multivesicular bodies) and in exosomes"
                     /translation="MAVEGGMKCVKFLLYVLLLAFCACAVGLIAVGVGAQLVLSQTIIQ
                     GATPGSLLPVVIIAVGVFLFLVAFVGCCGACKENYCLMITFAIFLSLIMLVEVAAAIAG
                     YVFRDKVMSEFNNNFRQQMENYPKNNHTASILDRMQADFKCCGAANYTDWEKIPSMSKN
                     RVPDSCCINVTVGCGINFNEKAIHKEGCVEKIGGWLRKNVLVVAAAALGIAFVEVLGIV
                     FACCLVKSIRSGYEVM"
     polyA_signal    1610..1731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1647..1666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1701..1720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1738..2193)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1875..1896)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2087..2106
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2220..2324
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2326..2683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2326..2517
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2349..2369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2534..2669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2718..3512
                     /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             2718..3512
                     /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(2772..2791)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3382..3401
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3700..3719)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3744..3791
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4120..4708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4609..4628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4770..4899
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        join(4923..5278,1..24)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
       61 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      121 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      181 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      241 cgtcagatcc gctagccacc atgaaatcaa ttcgctgtaa aaactgcaac aaactgttat
      301 ttaaggcgga ttcctttgat cacattgaaa tcaggtgtcc gcgttgcaaa cgtcacatca
      361 taatgctcaa cgcttgtgaa caccctacgg agaaacattg tgggaaaaga gaaaaaatca
      421 cgcattctga cgaaaccgtg cgttatggcg gtcacaactc cggaggtgga ggtggacaga
      481 gcccaggccc ggcagccatg gcggtggaag gaggaatgaa atgtgtgaag ttcttgctct
      541 acgtcctcct gctggccttt tgcgcctgtg cagtgggact gattgccgtg ggtgtcgggg
      601 cacagcttgt cctgagtcag accataatcc agggggctac ccctggctct ctgttgccag
      661 tggtcatcat cgcagtgggt gtcttcctct tcctggtggc ttttgtgggc tgctgcgggg
      721 cctgcaagga gaactattgt cttatgatca cgtttgccat ctttctgtct cttatcatgt
      781 tggtggaggt ggccgcagcc attgctggct atgtgtttag agataaggtg atgtcagagt
      841 ttaataacaa cttccggcag cagatggaga attacccgaa aaataaccac actgcttcga
      901 tcctggacag gatgcaggca gattttaagt gctgtggggc tgctaactac acagattggg
      961 agaaaatccc ttccatgtcg aagaaccgag tccccgactc ctgctgcatt aatgttactg
     1021 tgggctgtgg gattaatttc aacgagaagg cgatccataa ggagggctgt gtggagaaga
     1081 ttgggggctg gctgaggaaa aatgtgctgg tggtagctgc agcagccctt ggaattgctt
     1141 ttgtcgaggt tttgggaatt gtctttgcct gctgcctcgt gaagagtatc agaagtggct
     1201 acgaggtgat gtccggtgga ggtggatcca tggcttctaa ctttactcag ttcgttctcg
     1261 tcgacaatgg cggaactggc gacgtgatga aatcaattcg ctgtaaaaac tgcaacaaac
     1321 tgttatttaa ggcggattcc tttgatcaca ttgaaatcag gtgtccgcgt tgcaaacgtc
     1381 acatcataat gctcaacgct tgtgaacacc ctacggagaa acattgtggg aaaagagaaa
     1441 aaatcacgca ttctgacgaa accgtgcgtt attagagatc tgatccaccg gatctagata
     1501 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1561 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1621 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1681 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     1741 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1801 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1861 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1921 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1981 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2041 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2101 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2161 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     2221 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2281 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     2341 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2401 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2521 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2581 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2641 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     2701 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2761 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2821 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2881 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2941 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3001 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3061 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3121 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3181 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3241 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3301 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3361 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3421 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3481 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3541 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3601 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3661 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     3721 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     3781 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3841 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3901 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     3961 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4021 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4081 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4141 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4201 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4261 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4321 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4381 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4441 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4501 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4561 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4621 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4681 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4741 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
     4801 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     4861 gcagagaggg agtggccaac tccatcacta ggggttcctg cggcctctag actcgaggcg
     4921 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     4981 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     5041 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     5101 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     5161 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     5221 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctac
//
