LOCUS       sequence-319353-        6540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminally HiBiT-tagged VSV-G  under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6540)
  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 6540)
  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: pCMV-VSV-G-HiBiT
FEATURES             Location/Qualifiers
     source          1..6540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             join(1354..1401,1402..2886)
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     primer_bind     complement(3149..3168)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3214..3269
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3268..3287)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3627..3643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3627..3643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3640..3662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3651..3667
                     /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(3675..3681,3682..3699,3700..3705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3720..3741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3858..3875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4029..4617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4109..4128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4788..5579,5580..5648))
                     /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     5411..5430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5649..5753)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5779..6234)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5916..5937)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6128..6147
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6361..6383
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6375..6392
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6376..6392
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6402..6421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6402..6420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atccctcgag gaattctgac actatgaagt gccttttgta cttagccttt
     1381 ttattcattg gggtgaattg caagttcacc atagtttttc cacacaacca aaaaggaaac
     1441 tggaaaaatg ttccttctaa ttaccattat tgcccgtcaa gctcagattt aaattggcat
     1501 aatgacttaa taggcacagc cttacaagtc aaaatgccca agagtcacaa ggctattcaa
     1561 gcagacggtt ggatgtgtca tgcttccaaa tgggtcacta cttgtgattt ccgctggtat
     1621 ggaccgaagt atataacaca ttccatccga tccttcactc catctgtaga acaatgcaag
     1681 gaaagcattg aacaaacgaa acaaggaact tggctgaatc caggcttccc tcctcaaagt
     1741 tgtggatatg caactgtgac ggatgccgaa gcagtgattg tccaggtgac tcctcaccat
     1801 gtgctggttg atgaatacac aggagaatgg gttgattcac agttcatcaa cggaaaatgc
     1861 agcaattaca tatgccccac tgtccataac tctacaacct ggcattctga ctataaggtc
     1921 aaagggctat gtgattctaa cctcatttcc atggacatca ccttcttctc agaggacgga
     1981 gagctatcat ccctgggaaa ggagggcaca gggttcagaa gtaactactt tgcttatgaa
     2041 actggaggca aggcctgcaa aatgcaatac tgcaagcatt ggggagtcag actcccatca
     2101 ggtgtctggt tcgagatggc tgataaggat ctctttgctg cagccagatt ccctgaatgc
     2161 ccagaagggt caagtatctc tgctccatct cagacctcag tggatgtaag tctaattcag
     2221 gacgttgaga ggatcttgga ttattccctc tgccaagaaa cctggagcaa aatcagagcg
     2281 ggtcttccaa tctctccagt ggatctcagc tatcttgctc ctaaaaaccc aggaaccggt
     2341 cctgctttca ccataatcaa tggtacccta aaatactttg agaccagata catcagagtc
     2401 gatattgctg ctccaatcct ctcaagaatg gtcggaatga tcagtggaac taccacagaa
     2461 agggaactgt gggatgactg ggcaccatat gaagacgtgg aaattggacc caatggagtt
     2521 ctgaggacca gttcaggata taagtttcct ttatacatga ttggacatgg tatgttggac
     2581 tccgatcttc atcttagctc aaaggctcag gtgttcgaac atcctcacat tcaagacgct
     2641 gcttcgcaac ttcctgatga tgagagttta ttttttggtg atactgggct atccaaaaat
     2701 ccaatcgagc ttgtagaagg ttggttcagt agttggaaaa gctctattgc ctcttttttc
     2761 tttatcatag ggttaatcat tggactattc ttggttctcc gagttggtat ccatctttgc
     2821 attaaattaa agcacaccaa gaaaagacag atttatacag acatagagat gaaccgactt
     2881 ggaaaggtga gcggctggcg gctgttcaag aagattagct aactcaaatc ctgcacaaca
     2941 gattcttcat gtttggacca aatcaacttg tgataccatg ctcaaagagg cctcaattat
     3001 atttgagttt ttaattttta tgaaaaaaaa aaaaaaaaac ggaattcctc gagggatccg
     3061 tcgaggaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     3121 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     3181 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     3241 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     3301 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     3361 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     3421 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     3481 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     3541 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     3601 acggatcggc cgcaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     3661 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3721 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3781 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3841 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3901 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3961 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4021 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4081 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4141 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4201 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4261 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4321 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4381 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4441 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4501 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4561 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4621 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4681 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4741 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4801 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4861 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4921 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4981 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5041 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5101 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5161 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5221 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5281 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5341 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5401 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5461 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5521 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5581 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5641 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5761 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     5821 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     5881 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     5941 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6001 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6061 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     6121 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     6181 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     6241 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     6301 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     6361 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     6421 gcgaattgga gctccaccgc ggtggcggcc gctctagaga gcttggccca ttgcatacgt
     6481 tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt
//
