LOCUS       sequence-387085-        6465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpress CD63.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  TITLE     Bordey Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6465)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCAG-GFP-CD63
FEATURES             Location/Qualifiers
     source          1..6465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        372..648
                     /label=chicken beta-actin promoter
     intron          650..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1675..1694
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1741..1750
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1747..1749,1750..1752,1753..2463)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1747..1749,1750..1752,1753..2463)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1792..1813)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2053..2072)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2400..2421
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2539..3255
                     /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"
     primer_bind     complement(3473..3492)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3538..3593
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3592..3611)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3954..3970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3954..3970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3967..3989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3978..3994
                     /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(4002..4008,4009..4026,4027..4032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4047..4068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4127..4322
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4173..4308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4328..4462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4365..4384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4419..4438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4530..4547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4701..5289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4781..4800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5460..6251,6252..6320))
                     /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     6083..6102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6321..6425)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6456..6465,1..370)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      421 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      481 gggggggggg ggsgcgcgcc aggcggggsg gggcggggcg rggggcgggg cggggcgagg
      541 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      601 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg gagtcgctgc
      661 gacgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      721 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      781 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      841 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      901 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      961 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     1021 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1081 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     1141 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     1201 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     1261 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     1321 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1381 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     1441 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     1501 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgaggat gctaccggtc
     1741 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1801 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1861 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1921 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     1981 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2041 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2101 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2161 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2221 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2281 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2341 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2401 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2461 aagtccggcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccag
     2521 agcccaggcc cggcagccat ggcggtggaa ggaggaatga aatgtgtgaa gttcttgctc
     2581 tacgtcctcc tgctggcctt ttgcgcctgt gcagtgggac tgattgccgt gggtgtcggg
     2641 gcacagcttg tcctgagtca gaccataatc cagggggcta cccctggctc tctgttgcca
     2701 gtggtcatca tcgcagtggg tgtcttcctc ttcctggtgg cttttgtggg ctgctgcggg
     2761 gcctgcaagg agaactattg tcttatgatc acgtttgcca tctttctgtc tcttatcatg
     2821 ttggtggagg tggccgcagc cattgctggc tatgtgttta gagataaggt gatgtcagag
     2881 tttaataaca acttccggca gcagatggag aattacccga aaaataacca cactgcttcg
     2941 atcctggaca ggatgcaggc agattttaag tgctgtgggg ctgctaacta cacagattgg
     3001 gagaaaatcc cttccatgtc gaagaaccga gtccccgact cctgctgcat taatgttact
     3061 gtgggctgtg ggattaattt caacgagaag gcgatccata aggagggctg tgtggagaag
     3121 attgggggct ggctgaggaa aaatgtgctg gtggtagctg cagcagccct tggaattgct
     3181 tttgtcgagg ttttgggaat tgtctttgcc tgctgcctcg tgaagagtat cagaagtggc
     3241 tacgaggtga tgtagggatc caccggatct agataactga tcataatcag ccataccaca
     3301 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3361 aaaatgaatg caattgttgt tgttatcgcg gccgcactcc tcaggtgcag gctgcctatc
     3421 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     3481 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     3541 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     3601 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3661 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3721 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3781 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3841 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3901 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     3961 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4021 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4081 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     4141 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4201 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4261 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4321 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4441 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     4501 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4561 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4621 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5101 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5401 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5461 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5521 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5581 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5641 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5701 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5761 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5821 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5881 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5941 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6001 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6061 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6121 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6181 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6241 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6301 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6421 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgatt
//
