LOCUS       sequence-352551-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human CD63-FKBP-Myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6575)
  AUTHORS   Somiya M, Kuroda S
  TITLE     Engineering of Extracellular Vesicles for Small Molecule-Regulated 
            Cargo Loading and Cytoplasmic Delivery of Bioactive Proteins.
  JOURNAL   Mol Pharm. 2022 May 20. doi: 10.1021/acs.molpharmaceut.2c00192.
  PUBMED    35594496
REFERENCE   2  (bases 1 to 6575)
  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: "Mol Pharm. 
            2022 May 20. doi: 10.1021/acs.molpharmaceut.2c00192."
COMMENT     Sequence Label: pCMV-CD63-FKBP-Myc
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(118..137)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        309..688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        689..892
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     843..863
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     937..956
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        937..955
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1053..1766
                     /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"
     CDS             1791..2111
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2112..2141
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2166..2185)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     complement(2240..2257)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2246..2470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2516..2944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2603..2622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2813..2834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2953..2973)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2958..3287
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3138..3273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3200..3219
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3354..4148
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3408..3427)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4018..4037
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4322..4443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4359..4378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4413..4432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4492..4508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4492..4508)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4505..4527)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4516..4532
                     /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(4540..4546,4547..4564,4565..4570))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4585..4606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4723..4740)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4894..5482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4974..4993)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5653..6444,6445..6513))
                     /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     6276..6295
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6514..6575,1..43))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
       61 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
      121 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
      181 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
      241 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      301 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      361 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      421 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      481 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      541 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      601 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      661 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      721 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      781 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      841 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
      901 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      961 ccaagctggc tagcgtttaa acgggccctc tagaattatc gatccggagg tacccgggcg
     1021 gccgcgccac catgagccca ggcccggcag ccatggcggt ggaaggagga atgaaatgtg
     1081 tgaagttctt gctctacgtc ctcctgctgg ccttttgcgc ctgtgcagtg ggactgattg
     1141 ccgtgggtgt cggggcacag cttgtcctga gtcagaccat aatccagggg gctacccctg
     1201 gctctctgtt gccagtggtc atcatcgcag tgggtgtctt cctcttcctg gtggcttttg
     1261 tgggctgctg cggggcctgc aaggagaact attgtcttat gatcacgttt gccatctttc
     1321 tgtctcttat catgttggtg gaggtggccg cagccattgc tggctatgtg tttagagata
     1381 aggtgatgtc agagtttaat aacaacttcc ggcagcagat ggagaattac ccgaaaaata
     1441 accacactgc ttcgatcctg gacaggatgc aggcagattt taagtgctgt ggggctgcta
     1501 actacacaga ttgggagaaa atcccttcca tgtcgaagaa ccgagtcccc gactcctgct
     1561 gcattaatgt tactgtgggc tgtgggatta atttcaacga gaaggcgatc cataaggagg
     1621 gctgtgtgga gaagattggg ggctggctga ggaaaaatgt gctggtggta gctgcagcag
     1681 cccttggaat tgcttttgtc gaggttttgg gaattgtctt tgcctgctgc ctcgtgaaga
     1741 gtatcagaag tggctacgag gtgatgggag gcggaggcgg aggcggaggc ggagtgcagg
     1801 tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg
     1861 tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca
     1921 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg
     1981 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg
     2041 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc
     2101 taaaactgga agaacagaaa ctgattagcg aagaagatct gtaactcgag aagcttgcta
     2161 gcagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcactta
     2221 agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2281 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2341 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2401 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2461 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2881 attttgccga tttcggccta ttggttaaaa aataagctga tttaacaaaa atttaacgcg
     2941 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3301 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3361 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3421 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3481 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3541 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3601 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3661 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3721 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3781 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3841 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3901 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3961 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4021 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4081 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4141 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4201 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4261 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4321 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4381 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4441 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4501 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4561 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4621 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4681 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4741 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4801 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4861 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4921 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4981 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5041 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5101 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5161 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5221 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5281 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5341 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5401 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5461 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5521 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5581 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5641 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5701 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5761 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5821 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5881 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5941 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6001 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6061 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6121 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6181 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6241 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6301 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6361 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6421 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6481 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6541 catttatcag ggttattgtc tcatgagcgg ataca
//
