LOCUS       sequence-375170-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pVPSB backbone expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  AUTHORS   Yuan S, Peng L, Park JJ, Hu Y, Devarkar SC, Dong MB, Shen Q, Wu S, 
            Chen S, Lomakin IB, Xiong Y
  TITLE     Nonstructural Protein 1 of SARS-CoV-2 Is a Potent Pathogenicity 
            Factor Redirecting Host Protein Synthesis Machinery toward Viral 
            RNA.
  JOURNAL   Mol Cell. 2020 Dec 17;80(6):1055-1066.e6. doi: 
            10.1016/j.molcel.2020.10.034. Epub 2020 Oct 29.
  PUBMED    33188728
REFERENCE   2  (bases 1 to 5570)
  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"; doi: 
            "10.1016/j.molcel.2020.10.034"; journalName: "Mol Cell"; date: 
            "2020-12-17- 17"; volume: "80"; issue: "6"; pages: "1055-1066.e6"
COMMENT     Sequence Label: pVPSB_empty
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        581..792
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    824..872
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1161..1178)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1167..1391
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1437..1865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1524..1543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1734..1755
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1874..1894)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1879..2208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2059..2194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2121..2140
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2275..3069
                     /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(2329..2348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2939..2958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3243..3364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3280..3299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3334..3353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3413..3429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3413..3429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3426..3448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3437..3453
                     /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(3461..3467,3468..3485,3486..3491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3506..3527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3644..3661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3815..4403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3895..3914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4574..5365,5366..5434))
                     /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     5197..5216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5435..5539)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttctagac
      241 tatacagttg aagtcggaag tttacataca cttaagttgg agtcattaaa actcgttttt
      301 caactactcc acaaatttct tgttaacaaa caatagtttt ggcaagtcag ttaggacatc
      361 tactttgtgc atgacacaag tcatttttcc aacaattgtt tacagacaga ttatttcact
      421 tataattcac tgtatcacaa ttccagtggg tcagaagttt acatacacta agttgactgt
      481 gcctttaaac agcttggaaa attccagaaa atgatgtcat ggctttagag aggatcctag
      541 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
      601 acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg
      661 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
      721 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
      781 gccagaacac aggaccggtg gtacccgtac ggccaccctc gagaataaaa gatctttatt
      841 ttcattagat ctgtgtgttg gttttttgtg tgaattttga gtgtatgtaa acttctgacc
      901 cactgggaat gtgatgaaag aaataaaagc tgaaatgaat cattctctct actattattc
      961 tgatatttca cattcttaaa ataaagtggt gatcctaact gacctaagac agggaatttt
     1021 tactaggatt aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt ggctaaggtg
     1081 tatgtaaact tccgacttca actgtatagg catgcggtaa ccacgtgcgg accgaagctt
     1141 aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1201 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1261 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1321 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1381 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1441 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1501 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1561 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1621 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1681 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1741 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1801 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1861 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     1921 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     1981 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2041 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2101 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2161 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2221 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2281 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2341 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2401 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2461 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2521 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2581 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2641 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2701 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2761 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     2821 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     2881 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     2941 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3001 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3061 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3121 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3181 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3241 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3361 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3421 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3481 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3541 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3601 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4621 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4681 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4741 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4981 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
