LOCUS       sequence-325029-        5552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminally HiBiT-tagged human CD63 under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5552)
  AUTHORS   Somiya M, Kuroda S
  TITLE     Real-Time Luminescence Assay for Cytoplasmic Cargo Delivery of 
            Extracellular Vesicles.
  JOURNAL   Anal Chem. 2021 Mar 24. doi: 10.1021/acs.analchem.1c00339.
  PUBMED    33759512
REFERENCE   2  (bases 1 to 5552)
  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: "Anal Chem. 
            2021 Mar 24. doi: 10.1021/acs.analchem.1c00339."
COMMENT     Sequence Label: pCAG-HiBiT-CD63
FEATURES             Location/Qualifiers
     source          1..5552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLYRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        997..1376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1654..2670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2594..2616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2678..2697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2746..2755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2788..3501
                     /codon_start=1
                     /product="human CD63 tetraspanin"
                     /label=CD63
                     /note="enriched in intraluminal vesicles of late endosomes 
                     (multivesicular bodies) and in exosomes"
                     /translation="AVEGGMKCVKFLLYVLLLAFCACAVGLIAVGVGAQLVLSQTIIQG
                     ATPGSLLPVVIIAVGVFLFLVAFVGCCGACKENYCLMITFAIFLSLIMLVEVAAAIAGY
                     VFRDKVMSEFNNNFRQQMENYPKNNHTASILDRMQADFKCCGAANYTDWEKIPSMSKNR
                     VPDSCCINVTVGCGINFNEKAIHKEGCVEKIGGWLRKNVLVVAAAALGIAFVEVLGIVF
                     ACCLVKSIRSGYEVM"
     primer_bind     complement(3587..3606)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3652..3707
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3706..3725)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4068..4084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4068..4084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4081..4103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4092..4108
                     /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(4116..4122,4123..4140,4141..4146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4161..4182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      4267..4401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4421..4555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4458..4477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4512..4531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4623..4640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4794..5382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4874..4893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac ggtataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt
     1021 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     1081 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     1141 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     1201 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     1261 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1321 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     1381 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     1441 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggsgc
     1501 gcgccrggsg gggggggggg gggsgggggg gggggsgggg sgrggsggag aggtgcggcg
     1561 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     1621 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
     1681 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     1741 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
     1801 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
     1861 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg
     1921 tgcggctccg cgctacccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     1981 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
     2041 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
     2101 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
     2161 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
     2221 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
     2281 gggnaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
     2341 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     2401 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     2461 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     2521 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
     2581 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
     2641 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     2701 tctcatcatt ttggcaaaga attgattaat tcgagcgaac gcgtcgccac catggtgagc
     2761 ggctggcggc tgttcaagaa gattagcgcg gtggaaggag gaatgaaatg tgtgaagttc
     2821 ttgctctacg tcctcctgct ggccttttgc gcctgtgcag tgggactgat tgccgtgggt
     2881 gtcggggcac agcttgtcct gagtcagacc ataatccagg gggctacccc tggctctctg
     2941 ttgccagtgg tcatcatcgc agtgggtgtc ttcctcttcc tggtggcttt tgtgggctgc
     3001 tgcggggcct gcaaggagaa ctattgtctt atgatcacgt ttgccatctt tctgtctctt
     3061 atcatgttgg tggaggtggc cgcagccatt gctggctatg tgtttagaga taaggtgatg
     3121 tcagagttta ataacaactt ccggcagcag atggagaatt acccgaaaaa taaccacact
     3181 gcttcgatcc tggacaggat gcaggcagat tttaagtgct gtggggctgc taactacaca
     3241 gattgggaga aaatcccttc catgtcgaag aaccgagtcc ccgactcctg ctgcattaat
     3301 gttactgtgg gctgtgggat taatttcaac gagaaggcga tccataagga gggctgtgtg
     3361 gagaagattg ggggctggct gaggaaaaat gtgctggtgg tagctgcagc agcccttgga
     3421 attgcttttg tcgaggtttt gggaattgtc tttgcctgct gcctcgtgaa gagtatcaga
     3481 agtggctacg aggtgatgta agcggccgca ctcctcaggt gcaggctgcc tatcagaagg
     3541 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     3601 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3661 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3721 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3781 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3841 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3901 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3961 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     4021 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     4081 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4141 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4201 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     4261 ccatagtccc gctcctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4321 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4381 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     4441 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4501 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     4561 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4621 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4681 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5521 taaagtatat atgagtaaac ttggtctgac ag
//
