LOCUS       sequence-227501-        6406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is an envelope gene encoding plasmid to make retrogradely 
            lentiviral vector. If you use this plasmid, you can make type E 
            (P440E) envelope coating virus particle with NeuRet..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  AUTHORS   Kato S, Sugawara M, Kobayashi K, Kimura K, Inoue KI, Takada M, 
            Kobayashi K
  TITLE     Enhancement of the transduction efficiency of a lentiviral vector 
            for neuron-specific retrograde gene delivery through the point 
            mutation of fusion glycoprotein type E.
  JOURNAL   J Neurosci Methods. 2018 Oct 19;311:147-155. doi: 
            10.1016/j.jneumeth.2018.10.023.
  PUBMED    30347222
REFERENCE   2  (bases 1 to 6406)
  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: "J Neurosci 
            Methods."; date: "2018-10-19"; volume: "311:147-155. doi"; pages: " 
            10.1016/j.jneumeth.2018.10.023"
COMMENT     Sequence Label: pCAGGS-FuG-E (P440E)
FEATURES             Location/Qualifiers
     source          1..6406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3224..3256
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     primer_bind     complement(3430..3449)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3495..3550
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3549..3568)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3909..3925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3909..3925)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3922..3944)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3933..3949
                     /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(3957..3963,3964..3981,3982..3987))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4002..4023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4082..4277
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4128..4263
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4283..4417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4320..4339)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4374..4393
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4485..4502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4656..5244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4736..4755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5415..6206,6207..6275))
                     /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     6038..6057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6276..6380)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      541 ggcggggcga ggcggaaagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgcccttagg
     1741 aaagatggtt ccgcaggttc ttttgtttgt actccttctg ggtttttcgt tgtgtttcgg
     1801 gaagttcccc atttacacga taccagacga acttggtccc tggagcccta ttgacataca
     1861 ccatctcagc tgtccaaata acctggttgt ggaggatgaa ggatgtacca acctgtccga
     1921 gttctcctac atggaactca aagtgggata catctcagcc atcaaagtga acgggttcac
     1981 ttgcacaggt gttgtgacag aggcagagac ctacaccaac tttgttggtt atgtcacaac
     2041 cacattcaag agaaagcatt tccgccccac cccagacgca tgtagagccg cgtataactg
     2101 gaagatggcc ggtgacccca gatatgaaga gtccctacac aatccatacc ccgactacca
     2161 ctggcttcga actgtaagaa ccaccaaaga gtccctcatt atcatatccc caagtgtgac
     2221 agatttggac ccatatgaca aatcccttca ctcaagggtc ttccctggcg gaaagtgctc
     2281 aggaataacg gtgtcctcta cctactgctc aactaaccat gattacacca tttggatgcc
     2341 cgagaatccg agaccaagga caccttgtga catttttacc aatagcagag ggaagagagc
     2401 atccaacggg aacaagactt gcggctttgt ggatgaaaga ggcctgtata agtctctaaa
     2461 aggagcatgc aggctcaagt tatgtggagt tcttggactt agacttatgg atggaacatg
     2521 ggtcgcgatg caaacatcag atgagaccaa atggtgccct ccagatcagt tggtgaattt
     2581 gcacgacttt cgctcagacg agatcgagca tctcgttgtg gaggagttag ttaagaaaag
     2641 agaggaatgt ctggatgcat tagagtccat catgaccacc aagtcagtaa gtttcagacg
     2701 tctcagtcac ctgagaaaac ttgtcccagg gtttggaaaa gcatatacca tattcaacaa
     2761 aaccttgatg gaggctgatg ctcactacaa gtcagtccgg acctggaatg agatcatccc
     2821 ctcaaaaggg tgtttgaaag ttggaggaag gtgccatcct catgtgaacg gggtgttttt
     2881 caatggtata atattagggc ctgacgacca tgtcctaatc ccagagatgc aatcatccct
     2941 cctccagcaa catatggagt tgttggaatc ttcagttatc cccctgatgc accccctggc
     3001 agacccttct acagttttca aagaaggtga tgaggctgag gattttgttg aagttcacct
     3061 cgagaaaaat ccaatcgagc ttgtagaagg ttggttcagt agttggaaaa gctctattgc
     3121 ctcttttttc tttatcatag ggttaatcat tggactattc ttggttctcc gagttggtat
     3181 ccatctttgc attaaattaa agcacaccaa gaaaagacag atttatacag acatagagat
     3241 gaaccgactt ggaaagtaac tcaaatcctg cacaacagat tcttcatgtt tggaccaaat
     3301 caacttgtga taccatgctc aaagaggcct caattatatt tgagttttta atttttatga
     3361 aaaaaaaaaa aaaaaacgga attcctgcag cccccaacaa ccggtacctc tagaactata
     3421 gctagcagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     3481 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3541 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     3601 tttggtttag agtttggcaa catatgccat atgctggctg ccatgaacaa aggtggctat
     3661 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     3721 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     3781 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     3841 cagtcatagc tgtccctctt ctcttatgaa gatccctcga cctgcagccc aagcttggcg
     3901 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3961 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4021 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     4081 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4141 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4201 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4261 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     4321 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4381 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     4441 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4501 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4561 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4621 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4681 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4741 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4801 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     4861 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4921 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4981 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5041 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5101 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5161 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5221 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5281 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5341 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5401 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5461 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5521 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5581 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5641 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5701 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5761 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5821 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5881 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5941 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6001 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6061 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6121 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6181 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6241 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6301 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6361 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
