LOCUS       sequence-459826-        6691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding a codon-optimized N gene from 
            vesicular stomatitis virus, strain Indiana..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6691)
  AUTHORS   Marques MC, Andreu-Moreno I, Sanjuan R, Elena SF, Geller R
  TITLE     An efficient plasmid-based system for the recovery of recombinant 
            vesicular stomatitis virus encoding foreign glycoproteins.
  JOURNAL   Sci Rep. 2024 Jun 25;14(1):14644. doi: 10.1038/s41598-024-65384-8.
  PUBMED    38918479
REFERENCE   2  (bases 1 to 6691)
  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: "Sci Rep."; 
            date: "2024-06-25"; volume: "14(1):14644. doi"; pages: " 
            10.1038/s41598-024-65384-8"
COMMENT     Sequence Label: pCMV-Nopt
FEATURES             Location/Qualifiers
     source          1..6691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    47..168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(58..77)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     112..131
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      354..809
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(441..460)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     651..672
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        926..1283
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        926..1117
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(949..969)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1134..1269
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1334..2128
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1388..1407)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1998..2017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2192..2240
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2400..2422
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2460..2478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2546..2650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2651..2719,2720..3511)
                     /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     complement(2869..2888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3682..4270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4171..4190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4481..4860
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4861..5072
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5023..5043
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5069..5093
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5233..5365
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5348..5367
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gcggccgctt ccctttagtg agggttaatg cttcgagcag acatgataag atacattgat
       61 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
      121 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
      181 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
      241 aacctctaca aatgtggtaa aatccgataa ggatcgatcc gggctggcgt aatagcgaag
      301 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
      361 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      421 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      481 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      541 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
      601 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      661 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      721 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      781 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
      841 tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg
      901 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
      961 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     1021 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1081 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     1141 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     1201 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     1261 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc tcgaacttaa
     1321 ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     1381 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     1441 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     1501 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     1561 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     1621 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     1681 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     1741 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     1801 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     1861 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     1921 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     1981 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     2041 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     2101 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     2161 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     2221 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgcgtatggt gcactctcag
     2281 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     2341 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2401 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2461 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2521 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2581 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2641 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2701 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2761 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2821 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2881 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2941 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3001 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3061 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3121 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3181 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3241 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3301 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3361 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3421 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3481 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3541 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3601 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3661 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3721 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3781 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3841 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3901 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3961 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4021 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4081 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4141 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4201 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4261 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4321 tgctcacatg gctcgacaga tcttcaatat tggccattag ccatattatt cattggttat
     4381 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     4441 tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt
     4501 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4561 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4621 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4681 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4741 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4801 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4861 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
     4921 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     4981 tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc
     5041 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
     5101 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa
     5161 cagtctcgaa cttaagctgc agtgactctc ttaaggtagc cttgcagaag ttggtcgtga
     5221 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
     5281 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
     5341 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
     5401 tagagtactt aagctagcca ccatgagcgt gaccgtgaaa agaatcatcg ataataccgt
     5461 ggtggtcccc aagctgcccg ccaacgagga ccccgtggaa tatcctgccg actacttcag
     5521 aaaatccaag gaaatcccac tgtacatcaa caccacaaag agcctgtctg atctgcgcgg
     5581 ttatgtgtac cagggcctta agagcggcaa cgtgtctatc atccacgtga acagctacct
     5641 gtacggcgcc ctgaaggata tcagaggaaa actggataag gactggagca gcttcggcat
     5701 caatatcggc aaggctggcg acaccatcgg aatcttcgac ctcgtgtccc tgaaggccct
     5761 ggacggggtg ctgcctgatg gcgtgtccga cgccagcaga acctctgccg acgacaagtg
     5821 gctgccactg tacctgctgg gcctgtacag agtgggcaga acacagatgc ctgaataccg
     5881 gaagaaactg atggacggcc tgaccaatca atgtaaaatg atcaatgagc agtttgagcc
     5941 tctggtgccc gagggccggg acatctttga tgtgtgggga aacgacagca actacaccaa
     6001 gatcgtggcc gccgtggaca tgttcttcca catgtttaag aagcacgagt gcgccagctt
     6061 ccggtacggc accattgtga gcagattcaa ggactgcgcc gctctggcca ccttcggcca
     6121 cctgtgcaag atcaccggca tgagcaccga ggacgtgaca acctggatcc tgaacagaga
     6181 agtggccgac gagatggtcc agatgatgct gcctggccag gagatcgaca aggccgacag
     6241 ctacatgccc tacctgatcg acttcggcct atcttctaag agcccataca gcagcgtgaa
     6301 aaaccccgct tttcatttct ggggccagct gacagccctg ctgctgaggt ccacaagagc
     6361 cagaaacgcc agacagcctg acgacatcga gtacacaagc ctgaccaccg ccggcctgct
     6421 gtatgcatac gccgtcggat ctagcgccga tctggctcag cagttctgcg tgggcgataa
     6481 caagtacaca cctgacgata gcacaggcgg cctgaccacc aacgcccctc ctcagggcag
     6541 agatgttgtg gaatggctgg gctggttcga ggatcagaac cggaaaccta cccctgacat
     6601 gatgcaatac gctaagcggg ccgtgatgag cctgcaggga ctgcgggaaa agactattgg
     6661 aaagtacgcc aagagcgagt tcgataagtg a
//
