LOCUS       sequence-292475-        6720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6720)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6720)
  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"
COMMENT     Sequence Label: pGBW-m4046218
FEATURES             Location/Qualifiers
     source          1..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..493
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    52..169
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     464..487
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2416..2433
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2434..2457
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2502..2520)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2871..3674
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      3757..4739
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4817..4846
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4868..5455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5467..5494)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5518..5709)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5710..6501,6502..6570))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6333..6352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6571..6673)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6674..6705)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 caggtatgat ttaaatggtc agttagagcg gccgcttgct atattgaagt acggattaga
       61 agccgccgag cgggcgacag ccctccgacg gaagactctc ctccgtgcgt cctcgtcttc
      121 accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga
      181 ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac
      241 aaaccttcaa attaacgaat caaattaaca accataggat gataatgcga ttagtttttt
      301 agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat
      361 ataaatggaa aagctgcata accactttaa ctaatacttt caacattttc agtttgtatt
      421 acttcttatt caaatgtcat aaaagtatca acaaaaaatt gttaatatac ctctatactt
      481 taacgtcaag gagaaaaaac tataatgtac actcgctatg tcgataacaa cttctgtggc
      541 cctgatggct accctcttga gtgcattaaa gaccttctag cacgtgctgg taaagcttca
      601 tgcactttgt ccgaacaact ggactttatt gacactaaga ggggtgtata ctgctgccgt
      661 gaacatgagc atgaaattgc ttggtacacg gaacgttctg aaaagagcta tgaattgcag
      721 acaccttttg aaattaaatt ggcaaagaaa tttgacacct tcaatgggga atgtccaaat
      781 tttgtatttc ccttaaattc cataatcaag actattcaac caagggttga aaagaaaaag
      841 cttgatggct ttatgggtag aattcgatct gtctatccag ttgcgtcacc aaatgaatgc
      901 aaccaaatgt gcctttcaac tctcatgaag tgtgatcatt gtggtgaaac ttcatggcag
      961 acgggcgatt ttgttaaagc cacttgcgaa ttttgtggca ctgagaattt gactaaagaa
     1021 ggtgccacta cttgtggtta cttaccccaa aatgctgttg ttaaaattta ttgtccagca
     1081 tgtcacaatt cagaagtagg acctgagcat agtcttgccg aataccataa tgaatctggc
     1141 ttgaaaacca ttcttcgtaa gggtggtcgc actattgcct ttggaggctg tgtgttctct
     1201 tatgttggtt gccataacaa gtgtgcctat tgggttccac gtgctagcgc taacataggt
     1261 tgtaaccata caggtgttgt tggagaaggt tccgaaggtc ttaatgacaa ccttcttgaa
     1321 atactccaaa aagagaaagt caacatcaat attgttggtg actttaaact taatgaagag
     1381 atcgccatta ttttggcatc tttttctgct tccacaagtg cttttgtgga aactgtgaaa
     1441 ggtttggatt ataaagcatt caaacaaatt gttgaatcct gtggtaattt taaagttaca
     1501 aaaggaaaag ctaaaaaagg tgcctggaat attggtgaac agaaatcaat actgagtcct
     1561 ctttatgcat ttgcatcaga ggctgctcgt gttgtacgat caattttctc ccgcactctt
     1621 gaaactgctc aaaattctgt gcgtgtttta cagaaggccg ctataacaat actagatgga
     1681 atttcacagt attcactgag actcattgat gctatgatgt tcacatctga tttggctact
     1741 aacaatctag ttgtaatggc ctacattaca ggtggtgttg ttcagttgac ttcgcagtgg
     1801 ctaactaaca tctttggcac tgtttatgaa aaactcaaac ccgtccttga ttggcttgaa
     1861 gagaagttta aggaaggtgt agagtttctt agagacggtt gggaaattgt taaatttatc
     1921 tcaacctgtg cttgtgaaat tgtcggtgga caaattgtca cctgtgcaaa ggaaattaag
     1981 gagagtgttc agacattctt taagcttgta aataaatttt tggctttgtg tgctgactct
     2041 atcattattg gtggagctaa acttaaagcc ttgaatttag gtgaaacatt tgtcacgcac
     2101 tcaaagggat tgtacagaaa gtgtgttaaa tccagagaag aaactggcct actcatgcct
     2161 ctaaaagccc caaaagaaat tatcttctta gagggagaaa cacttcccac agaagtgtta
     2221 acagaggaag ttgtcttgaa aactggtgat ttacaaccat tagaacaacc tactagtgaa
     2281 gctgttgaag ctccattggt tggtacacca gtttgtatta acgggcttat gttgctcgaa
     2341 atcaaagaca cagaaaagta ctgtgccctt gcacctaata tgatggtaac aaacaatacc
     2401 ttcacactca aaggcctcgt gccgagaggt tcttggagcc atccgcagtt tgaaaaataa
     2461 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     2521 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     2581 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     2641 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     2701 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     2761 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     2821 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     2881 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     2941 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     3001 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     3061 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     3121 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     3181 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     3241 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     3301 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     3361 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     3421 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     3481 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     3541 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     3601 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     3661 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     3721 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     3781 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3841 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     3901 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     3961 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     4021 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     4081 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     4141 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     4201 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     4261 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     4321 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     4381 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     4441 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     4501 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     4561 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     4621 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     4681 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     4741 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     4801 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     4861 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4921 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     4981 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5041 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5101 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5161 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5221 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5281 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5341 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5401 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5461 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     5521 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5581 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5641 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5701 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     6601 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     6661 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
//
