LOCUS       sequence-298145-        6687 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6687)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6687)
  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-m4046247
FEATURES             Location/Qualifiers
     source          1..6687
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             2314..2331
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2332..2397
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2442..2460)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2811..3614
                     /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      3697..4679
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4757..4786
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4808..5395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5407..5434)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5458..5649)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5650..6441,6442..6510))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGXQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6273..6292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6511..6613)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6614..6645)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt
      301 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt
      361 agaaagaaag catagcaatc taatctaagt tttaattaca aaatgtacac tcgctatgtc
      421 gataacaact tctgtggccc tgatggctac cctcttgagt gcattaaaga ccttctagca
      481 cgtgctggta aagcttcatg cactttgtcc gaacaactgg actttattga cactaagagg
      541 ggtgtatact gctgccgtga acatgagcat gaaattgctt ggtacacgga acgttctgaa
      601 aagagctatg aattgcagac accttttgaa attaaattgg caaagaaatt tgacaccttc
      661 aatggggaat gtccaaattt tgtatttccc ttaaattcca taatcaagac tattcaacca
      721 agggttgaaa agaaaaagct tgatggcttt atgggtagaa ttcgatctgt ctatccagtt
      781 gcgtcaccaa atgaatgcaa ccaaatgtgc ctttcaactc tcatgaagtg tgatcattgt
      841 ggtgaaactt catggcagac gggcgatttt gttaaagcca cttgcgaatt ttgtggcact
      901 gagaatttga ctaaagaagg tgccactact tgtggttact taccccaaaa tgctgttgtt
      961 aaaatttatt gtccagcatg tcacaattca gaagtaggac ctgagcatag tcttgccgaa
     1021 taccataatg aatctggctt gaaaaccatt cttcgtaagg gtggtcgcac tattgccttt
     1081 ggaggctgtg tgttctctta tgttggttgc cataacaagt gtgcctattg ggttccacgt
     1141 gctagcgcta acataggttg taaccataca ggtgttgttg gagaaggttc cgaaggtctt
     1201 aatgacaacc ttcttgaaat actccaaaaa gagaaagtca acatcaatat tgttggtgac
     1261 tttaaactta atgaagagat cgccattatt ttggcatctt tttctgcttc cacaagtgct
     1321 tttgtggaaa ctgtgaaagg tttggattat aaagcattca aacaaattgt tgaatcctgt
     1381 ggtaatttta aagttacaaa aggaaaagct aaaaaaggtg cctggaatat tggtgaacag
     1441 aaatcaatac tgagtcctct ttatgcattt gcatcagagg ctgctcgtgt tgtacgatca
     1501 attttctccc gcactcttga aactgctcaa aattctgtgc gtgttttaca gaaggccgct
     1561 ataacaatac tagatggaat ttcacagtat tcactgagac tcattgatgc tatgatgttc
     1621 acatctgatt tggctactaa caatctagtt gtaatggcct acattacagg tggtgttgtt
     1681 cagttgactt cgcagtggct aactaacatc tttggcactg tttatgaaaa actcaaaccc
     1741 gtccttgatt ggcttgaaga gaagtttaag gaaggtgtag agtttcttag agacggttgg
     1801 gaaattgtta aatttatctc aacctgtgct tgtgaaattg tcggtggaca aattgtcacc
     1861 tgtgcaaagg aaattaagga gagtgttcag acattcttta agcttgtaaa taaatttttg
     1921 gctttgtgtg ctgactctat cattattggt ggagctaaac ttaaagcctt gaatttaggt
     1981 gaaacatttg tcacgcactc aaagggattg tacagaaagt gtgttaaatc cagagaagaa
     2041 actggcctac tcatgcctct aaaagcccca aaagaaatta tcttcttaga gggagaaaca
     2101 cttcccacag aagtgttaac agaggaagtt gtcttgaaaa ctggtgattt acaaccatta
     2161 gaacaaccta ctagtgaagc tgttgaagct ccattggttg gtacaccagt ttgtattaac
     2221 gggcttatgt tgctcgaaat caaagacaca gaaaagtact gtgcccttgc acctaatatg
     2281 atggtaacaa acaatacctt cacactcaaa ggcctcgtgc cgagaggttc tgattataaa
     2341 gatcatgatg gtgattataa agatcatgat attgattata aagatgatga tgataaataa
     2401 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     2461 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     2521 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     2581 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     2641 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     2701 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     2761 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     2821 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     2881 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     2941 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     3001 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     3061 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     3121 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     3181 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     3241 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     3301 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     3361 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     3421 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     3481 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     3541 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     3601 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     3661 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     3721 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3781 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     3841 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     3901 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     3961 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     4021 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     4081 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     4141 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     4201 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     4261 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     4321 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     4381 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     4441 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     4501 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     4561 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     4621 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     4681 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     4741 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     4801 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4861 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     4921 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4981 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5041 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5101 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5161 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5221 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5281 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5341 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5401 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     5461 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5521 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5581 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5641 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5701 ttcatccata gttgcctgmc tccccgtcgt gtagataact acgatgcggg agggcttacc
     5761 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5821 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     5881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     6121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     6181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6301 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     6361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6481 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     6541 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     6601 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     6661 caggtatgat ttaaatggtc agttaga
//
