LOCUS       sequence-375077-        7437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express SARS2-NSP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7437)
  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 7437)
  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: pVPSBA01-SARS-CoV2-nsp1
FEATURES             Location/Qualifiers
     source          1..7437
                     /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"
     regulatory      821..830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1385..1408
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1445..1468
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    join(1480..2053,2054..2056,2057..2065)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1646..1663)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1873..1892
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2085..2684
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2085..2104)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2581..2601
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2691..2739
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3028..3045)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3034..3258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3304..3732
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3391..3410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3601..3622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3741..3761)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3746..4075
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3926..4061
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3988..4007
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4142..4936
                     /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(4196..4215)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4806..4825
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5110..5231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5147..5166)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5201..5220
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5280..5296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5280..5296)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5293..5315)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5304..5320
                     /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(5328..5334,5335..5352,5353..5358))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5373..5394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5511..5528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5682..6270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5762..5781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6441..7232,7233..7301))
                     /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     7064..7083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7302..7406)
                     /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 ggccaccgcc gccaccatgg agagtcttgt
      841 acccggcttt aacgagaaaa cacatgttca attgtcattg cctgtgctgc aagtccggga
      901 cgtactggtt agggggttcg gagattcagt agaagaggtg ctgtctgagg cgaggcaaca
      961 tttgaaagac gggacgtgcg ggctggttga ggtggaaaag ggagtacttc cacagttgga
     1021 acaaccgtat gtttttatta agcggagtga tgccaggact gcgccgcacg gtcatgtcat
     1081 ggttgagctt gtcgcagagc ttgaaggcat tcaatatggg cgctccggtg aaactcttgg
     1141 agtacttgtg ccgcacgtag gcgaaattcc agtggcatat cggaaggtgc tgttgagaaa
     1201 aaatgggaac aagggcgcag ggggccacag ttatggagcc gacctcaaat cctttgacct
     1261 tggcgacgag ttggggacag atccctacga ggatttccag gaaaactgga atacgaaaca
     1321 tagcagtggc gttactcgcg agttgatgcg cgaactgaac ggcggcctcg aaggcggcgg
     1381 gggatggagc catccacaat tcgagaaagg cggtggttca ggaggaggta gcgggggtgg
     1441 atcatggtca catccgcagt ttgaaaagta aggatcccgc ccctctccct ccccccccct
     1501 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     1561 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     1621 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     1681 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     1741 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     1801 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     1861 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     1921 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     1981 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     2041 acacgatgat aagcttgcca caacccacaa ggagacgacc ttccatgacc gagtacaagc
     2101 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     2161 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     2221 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     2281 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     2341 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     2401 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     2461 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     2521 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     2581 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     2641 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctagctcgag aataaaagat
     2701 ctttattttc attagatctg tgtgttggtt ttttgtgtga attttgagtg tatgtaaact
     2761 tctgacccac tgggaatgtg atgaaagaaa taaaagctga aatgaatcat tctctctact
     2821 attattctga tatttcacat tcttaaaata aagtggtgat cctaactgac ctaagacagg
     2881 gaatttttac taggattaaa tgtcaggaat tgtgaaaaag tgagtttaaa tgtatttggc
     2941 taaggtgtat gtaaacttcc gacttcaact gtataggcat gcggtaacca cgtgcggacc
     3001 gaagcttaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3061 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3121 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3181 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3241 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3301 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3361 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3421 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3481 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3541 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3601 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3661 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3721 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3781 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3841 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3901 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3961 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4021 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4081 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4141 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4201 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4261 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4321 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4381 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4441 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4501 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4561 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4621 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4681 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4741 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4801 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4861 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4921 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4981 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5041 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5101 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5161 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5221 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5281 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5341 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5401 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5461 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5521 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5581 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5641 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5701 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5761 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5821 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5881 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5941 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6001 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6061 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6121 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6181 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6481 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6541 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6841 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7321 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7381 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
