LOCUS       sequence-403312-        6265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a VSV-G variant (K47Q R354A) defective for LDL-R receptor 
            binding (CAG promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6265)
  AUTHORS   Hamilton JR, Chen E, Perez BS, Sandoval Espinoza CR, Kang MH, 
            Trinidad M, Ngo W, Doudna JA
  TITLE     In vivo human T cell engineering with enveloped delivery vehicles.
  JOURNAL   Nat Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z.
  PUBMED    38212493
REFERENCE   2  (bases 1 to 6265)
  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 
            Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z."
COMMENT     Sequence Label: pJRH-1187 VSVGmut
FEATURES             Location/Qualifiers
     source          1..6265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..664
                     /label=chicken beta-actin promoter
     intron          665..1681
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1605..1627
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1689..1708
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1742..1789,1790..3277)
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPQSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTEAELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     primer_bind     complement(3287..3306)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3352..3407
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3406..3425)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3768..3784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3768..3784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3781..3803)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3792..3808
                     /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(3816..3822,3823..3840,3841..3846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3861..3882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3941..4136
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3987..4122
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4142..4276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4179..4198)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4233..4252
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4344..4361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4515..5103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4595..4614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5274..6065,6066..6134))
                     /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     5897..5916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6135..6239)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg ggggggggcg cgcgccaggs ggggcggggc ggggcgaggg
      541 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
      721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
      841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
      901 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
      961 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     1021 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     1081 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     1141 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     1201 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     1261 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     1321 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     1381 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     1441 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     1501 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     1561 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     1621 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     1681 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcgccac
     1741 catgaagtgc cttttgtact tagccttttt attcattggg gtgaattgca agttcaccat
     1801 agtttttcca cacaaccaaa aaggaaactg gaaaaatgtt ccttctaatt accattattg
     1861 cccgtcaagc tcagatttaa attggcataa tgacttaata ggcacagcct tacaagtcaa
     1921 aatgccccaa agtcacaagg ctattcaagc agacggttgg atgtgtcatg cttccaaatg
     1981 ggtcactact tgtgatttcc gctggtatgg accgaagtat ataacacatt ccatccgatc
     2041 cttcactcca tctgtagaac aatgcaagga aagcattgaa caaacgaaac aaggaacttg
     2101 gctgaatcca ggcttccctc ctcaaagttg tggatatgca actgtgacgg atgccgaagc
     2161 agtgattgtc caggtgactc ctcaccatgt gctggttgat gaatacacag gagaatgggt
     2221 tgattcacag ttcatcaacg gaaaatgcag caattacata tgccccactg tccataactc
     2281 tacaacctgg cattctgact ataaggtcaa agggctatgt gattctaacc tcatttccat
     2341 ggacatcacc ttcttctcag aggacggaga gctatcatcc ctgggaaagg agggcacagg
     2401 gttcagaagt aactactttg cttatgaaac tggaggcaag gcctgcaaaa tgcaatactg
     2461 caagcattgg ggagtcagac tcccatcagg tgtctggttc gagatggctg ataaggatct
     2521 ctttgctgca gccagattcc ctgaatgccc agaagggtca agtatctctg ctccatctca
     2581 gacctcagtg gatgtaagtc taattcagga cgttgagagg atcttggatt attccctctg
     2641 ccaagaaacc tggagcaaaa tcagagcggg tcttccaatc tctccagtgg atctcagcta
     2701 tcttgctcct aaaaacccag gaaccggtcc tgctttcacc ataatcaatg gtaccctaaa
     2761 atactttgag accagataca tcagagtcga tattgctgct ccaatcctct caagaatggt
     2821 cggaatgatc agtggaacta ccacagaagc tgaactgtgg gatgactggg caccatatga
     2881 agacgtggaa attggaccca atggagttct gaggaccagt tcaggatata agtttccttt
     2941 atacatgatt ggacatggta tgttggactc cgatcttcat cttagctcaa aggctcaggt
     3001 gttcgaacat cctcacattc aagacgctgc ttcgcaactt cctgatgatg agagtttatt
     3061 ttttggtgat actgggctat ccaaaaatcc aatcgagctt gtagaaggtt ggttcagtag
     3121 ttggaaaagc tctattgcct cttttttctt tatcataggg ttaatcattg gactattctt
     3181 ggttctccga gttggtatcc atctttgcat taaattaaag cacaccaaga aaagacagat
     3241 ttatacagac atagagatga accgacttgg aaagtaagct agcagatctt tttccctctg
     3301 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3361 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3421 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3481 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3541 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3601 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3661 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     3721 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     3781 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3841 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3901 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     3961 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4021 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4081 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4141 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4261 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4321 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4381 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4441 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4501 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4561 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4621 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4681 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4741 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4801 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4861 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4921 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4981 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5041 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5101 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5161 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5221 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5281 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5341 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5401 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5461 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5521 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5581 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5641 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5701 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5761 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5821 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5881 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5941 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6001 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6061 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6121 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6181 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6241 acatttcccc gaaaagtgcc acctg
//
