LOCUS       sequence-275569-        6507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VSV-G envelope protein for pseudotyping NanoMEDIC 
            particle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6507)
  AUTHORS   Gee P, Lung MSY, Okuzaki Y, Sasakawa N, Iguchi T, Makita Y, Hozumi 
            H, Miura Y, Yang LF, Iwasaki M, Wang XH, Waller MA, Shirai N, Abe 
            YO, Fujita Y, Watanabe K, Kagita A, Iwabuchi KA, Yasuda M, Xu H, 
            Noda T, Komano J, Sakurai H, Inukai N, Hotta A
  TITLE     Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and 
            sgRNA to induce therapeutic exon skipping.
  JOURNAL   Nat Commun. 2020 Mar 13;11(1):1334. doi: 10.1038/s41467-020-14957-y.
  PUBMED    32170079
REFERENCE   2  (bases 1 to 6507)
  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: "Nat 
            Commun."; date: "2020-03-13"; volume: "11(1):1334. doi"; pages: " 
            10.1038/s41467-020-14957-y"
COMMENT     Sequence Label: pVSV-G
FEATURES             Location/Qualifiers
     source          1..6507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          316..888
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             join(974..1021,1022..2509)
                     /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(2736..2755)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2801..2856
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2855..2874)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3214..3230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3214..3230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3227..3249)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3238..3254
                     /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(3262..3268,3269..3286,3287..3292))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3307..3328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3445..3462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3616..4204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3696..3715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4375..5166,5167..5235))
                     /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     4998..5017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5236..5340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5366..5821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5503..5524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5715..5734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5948..5970
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5962..5979
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5963..5979
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5989..6008
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5989..6007
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        6128..6507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gtcgaccgat
      301 cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta
      361 aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt
      421 cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga
      481 caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc
      541 ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt
      601 ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt
      661 tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc
      721 tggcgtggaa atattcttat tggtagaaac aactacaccc tggtcatcat cctgcctttc
      781 tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa
      841 ccgggcccct ctgctaacca tgttcatgcc ttcttctctt tcctacagct cctgggcaac
      901 gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tcctcgacgg atccctcgag
      961 gaattctgac actatgaagt gccttttgta cttagccttt ttattcattg gggtgaattg
     1021 caagttcacc atagtttttc cacacaacca aaaaggaaac tggaaaaatg ttccttctaa
     1081 ttaccattat tgcccgtcaa gctcagattt aaattggcat aatgacttaa taggcacagc
     1141 cttacaagtc aaaatgccca agagtcacaa ggctattcaa gcagacggtt ggatgtgtca
     1201 tgcttccaaa tgggtcacta cttgtgattt ccgctggtat ggaccgaagt atataacaca
     1261 ttccatccga tccttcactc catctgtaga acaatgcaag gaaagcattg aacaaacgaa
     1321 acaaggaact tggctgaatc caggcttccc tcctcaaagt tgtggatatg caactgtgac
     1381 ggatgccgaa gcagtgattg tccaggtgac tcctcaccat gtgctggttg atgaatacac
     1441 aggagaatgg gttgattcac agttcatcaa cggaaaatgc agcaattaca tatgccccac
     1501 tgtccataac tctacaacct ggcattctga ctataaggtc aaagggctat gtgattctaa
     1561 cctcatttcc atggacatca ccttcttctc agaggacgga gagctatcat ccctgggaaa
     1621 ggagggcaca gggttcagaa gtaactactt tgcttatgaa actggaggca aggcctgcaa
     1681 aatgcaatac tgcaagcatt ggggagtcag actcccatca ggtgtctggt tcgagatggc
     1741 tgataaggat ctctttgctg cagccagatt ccctgaatgc ccagaagggt caagtatctc
     1801 tgctccatct cagacctcag tggatgtaag tctaattcag gacgttgaga ggatcttgga
     1861 ttattccctc tgccaagaaa cctggagcaa aatcagagcg ggtcttccaa tctctccagt
     1921 ggatctcagc tatcttgctc ctaaaaaccc aggaaccggt cctgctttca ccataatcaa
     1981 tggtacccta aaatactttg agaccagata catcagagtc gatattgctg ctccaatcct
     2041 ctcaagaatg gtcggaatga tcagtggaac taccacagaa agggaactgt gggatgactg
     2101 ggcaccatat gaagacgtgg aaattggacc caatggagtt ctgaggacca gttcaggata
     2161 taagtttcct ttatacatga ttggacatgg tatgttggac tccgatcttc atcttagctc
     2221 aaaggctcag gtgttcgaac atcctcacat tcaagacgct gcttcgcaac ttcctgatga
     2281 tgagagttta ttttttggtg atactgggct atccaaaaat ccaatcgagc ttgtagaagg
     2341 ttggttcagt agttggaaaa gctctattgc ctcttttttc tttatcatag ggttaatcat
     2401 tggactattc ttggttctcc gagttggtat ccatctttgc attaaattaa agcacaccaa
     2461 gaaaagacag atttatacag acatagagat gaaccgactt ggaaagtaac tcaaatcctg
     2521 cacaacagat tcttcatgtt tggaccaaat caacttgtga taccatgctc aaagaggcct
     2581 caattatatt tgagttttta atttttatga aaaaaaaaaa aaaaaacgga attcctcgag
     2641 ggatccgtcg aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     2701 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     2761 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     2821 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2881 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     2941 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     3001 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     3061 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     3121 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     3181 tccctcgacg gatcggccgc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3241 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3301 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3361 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3421 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3481 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3541 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3781 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4021 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4141 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4201 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4261 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4321 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4381 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4441 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4501 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4561 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4621 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4681 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4741 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4801 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4861 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4921 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4981 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5041 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5101 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5161 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5221 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5281 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5341 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5401 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5461 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5521 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5581 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5641 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5701 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5761 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5821 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5881 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     5941 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     6001 ctatagggcg aattggagct ccaccgcggt ggcggccgct ctagagagct tggcccattg
     6061 catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc aacattaccg
     6121 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     6181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6361 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6421 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     6481 tacgtattag tcatcgctat taccatg
//
