LOCUS       sequence-298483-        6613 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 6613)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6613)
  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-m4046694
FEATURES             Location/Qualifiers
     source          1..6613
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(2368..2386)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2737..3540
                     /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="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKXXL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      3623..4605
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4683..4712
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4734..5321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5333..5360)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5384..5575)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5576..6367,6368..6436))
                     /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     6199..6218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6437..6539)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6540..6571)
                     /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 caataaaatt tcttgaaaat
      241 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
      301 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
      361 cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca aaatgtacac
      421 tcgctatgtc gataacaact tctgtggccc tgatggctac cctcttgagt gcattaaaga
      481 ccttctagca cgtgctggta aagcttcatg cactttgtcc gaacaactgg actttattga
      541 cactaagagg ggtgtatact gctgccgtga acatgagcat gaaattgctt ggtacacgga
      601 acgttctgaa aagagctatg aattgcagac accttttgaa attaaattgg caaagaaatt
      661 tgacaccttc aatggggaat gtccaaattt tgtatttccc ttaaattcca taatcaagac
      721 tattcaacca agggttgaaa agaaaaagct tgatggcttt atgggtagaa ttcgatctgt
      781 ctatccagtt gcgtcaccaa atgaatgcaa ccaaatgtgc ctttcaactc tcatgaagtg
      841 tgatcattgt ggtgaaactt catggcagac gggcgatttt gttaaagcca cttgcgaatt
      901 ttgtggcact gagaatttga ctaaagaagg tgccactact tgtggttact taccccaaaa
      961 tgctgttgtt aaaatttatt gtccagcatg tcacaattca gaagtaggac ctgagcatag
     1021 tcttgccgaa taccataatg aatctggctt gaaaaccatt cttcgtaagg gtggtcgcac
     1081 tattgccttt ggaggctgtg tgttctctta tgttggttgc cataacaagt gtgcctattg
     1141 ggttccacgt gctagcgcta acataggttg taaccataca ggtgttgttg gagaaggttc
     1201 cgaaggtctt aatgacaacc ttcttgaaat actccaaaaa gagaaagtca acatcaatat
     1261 tgttggtgac tttaaactta atgaagagat cgccattatt ttggcatctt tttctgcttc
     1321 cacaagtgct tttgtggaaa ctgtgaaagg tttggattat aaagcattca aacaaattgt
     1381 tgaatcctgt ggtaatttta aagttacaaa aggaaaagct aaaaaaggtg cctggaatat
     1441 tggtgaacag aaatcaatac tgagtcctct ttatgcattt gcatcagagg ctgctcgtgt
     1501 tgtacgatca attttctccc gcactcttga aactgctcaa aattctgtgc gtgttttaca
     1561 gaaggccgct ataacaatac tagatggaat ttcacagtat tcactgagac tcattgatgc
     1621 tatgatgttc acatctgatt tggctactaa caatctagtt gtaatggcct acattacagg
     1681 tggtgttgtt cagttgactt cgcagtggct aactaacatc tttggcactg tttatgaaaa
     1741 actcaaaccc gtccttgatt ggcttgaaga gaagtttaag gaaggtgtag agtttcttag
     1801 agacggttgg gaaattgtta aatttatctc aacctgtgct tgtgaaattg tcggtggaca
     1861 aattgtcacc tgtgcaaagg aaattaagga gagtgttcag acattcttta agcttgtaaa
     1921 taaatttttg gctttgtgtg ctgactctat cattattggt ggagctaaac ttaaagcctt
     1981 gaatttaggt gaaacatttg tcacgcactc aaagggattg tacagaaagt gtgttaaatc
     2041 cagagaagaa actggcctac tcatgcctct aaaagcccca aaagaaatta tcttcttaga
     2101 gggagaaaca cttcccacag aagtgttaac agaggaagtt gtcttgaaaa ctggtgattt
     2161 acaaccatta gaacaaccta ctagtgaagc tgttgaagct ccattggttg gtacaccagt
     2221 ttgtattaac gggcttatgt tgctcgaaat caaagacaca gaaaagtact gtgcccttgc
     2281 acctaatatg atggtaacaa acaatacctt cacactcaaa ggctaaacag gccccttttc
     2341 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2401 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2461 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2521 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     2581 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     2641 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     2701 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     2761 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     2821 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggnnntact ggagttagtt
     2881 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     2941 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     3001 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     3061 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     3121 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     3181 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3241 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     3301 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     3361 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     3421 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     3481 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     3541 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     3601 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     3661 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3721 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     3781 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3841 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     3901 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     3961 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     4021 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     4081 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     4141 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     4201 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4261 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     4321 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     4381 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     4441 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     4501 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     4561 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     4621 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     4681 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     4741 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4801 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     4861 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4921 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4981 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5041 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5101 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5161 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5221 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     5281 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     5341 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     5401 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     5461 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     5521 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     5581 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5641 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     5701 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5761 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     5821 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5881 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5941 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6001 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     6061 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     6121 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6181 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6241 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     6301 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6361 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6421 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     6481 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     6541 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     6601 atggtcagtt aga
//
