LOCUS       sequence-299951-        6673 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 6673)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6673)
  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-m4046184
FEATURES             Location/Qualifiers
     source          1..6673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2342..2359
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2360..2383
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2428..2446)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2797..3600
                     /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      3683..4665
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4743..4772
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4794..5381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5393..5420)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5444..5635)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5636..6427,6428..6496))
                     /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     6259..6278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6497..6599)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6600..6631)
                     /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 ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtacactc gctatgtcga taacaacttc tgtggccctg atggctaccc
      481 tcttgagtgc attaaagacc ttctagcacg tgctggtaaa gcttcatgca ctttgtccga
      541 acaactggac tttattgaca ctaagagggg tgtatactgc tgccgtgaac atgagcatga
      601 aattgcttgg tacacggaac gttctgaaaa gagctatgaa ttgcagacac cttttgaaat
      661 taaattggca aagaaatttg acaccttcaa tggggaatgt ccaaattttg tatttccctt
      721 aaattccata atcaagacta ttcaaccaag ggttgaaaag aaaaagcttg atggctttat
      781 gggtagaatt cgatctgtct atccagttgc gtcaccaaat gaatgcaacc aaatgtgcct
      841 ttcaactctc atgaagtgtg atcattgtgg tgaaacttca tggcagacgg gcgattttgt
      901 taaagccact tgcgaatttt gtggcactga gaatttgact aaagaaggtg ccactacttg
      961 tggttactta ccccaaaatg ctgttgttaa aatttattgt ccagcatgtc acaattcaga
     1021 agtaggacct gagcatagtc ttgccgaata ccataatgaa tctggcttga aaaccattct
     1081 tcgtaagggt ggtcgcacta ttgcctttgg aggctgtgtg ttctcttatg ttggttgcca
     1141 taacaagtgt gcctattggg ttccacgtgc tagcgctaac ataggttgta accatacagg
     1201 tgttgttgga gaaggttccg aaggtcttaa tgacaacctt cttgaaatac tccaaaaaga
     1261 gaaagtcaac atcaatattg ttggtgactt taaacttaat gaagagatcg ccattatttt
     1321 ggcatctttt tctgcttcca caagtgcttt tgtggaaact gtgaaaggtt tggattataa
     1381 agcattcaaa caaattgttg aatcctgtgg taattttaaa gttacaaaag gaaaagctaa
     1441 aaaaggtgcc tggaatattg gtgaacagaa atcaatactg agtcctcttt atgcatttgc
     1501 atcagaggct gctcgtgttg tacgatcaat tttctcccgc actcttgaaa ctgctcaaaa
     1561 ttctgtgcgt gttttacaga aggccgctat aacaatacta gatggaattt cacagtattc
     1621 actgagactc attgatgcta tgatgttcac atctgatttg gctactaaca atctagttgt
     1681 aatggcctac attacaggtg gtgttgttca gttgacttcg cagtggctaa ctaacatctt
     1741 tggcactgtt tatgaaaaac tcaaacccgt ccttgattgg cttgaagaga agtttaagga
     1801 aggtgtagag tttcttagag acggttggga aattgttaaa tttatctcaa cctgtgcttg
     1861 tgaaattgtc ggtggacaaa ttgtcacctg tgcaaaggaa attaaggaga gtgttcagac
     1921 attctttaag cttgtaaata aatttttggc tttgtgtgct gactctatca ttattggtgg
     1981 agctaaactt aaagccttga atttaggtga aacatttgtc acgcactcaa agggattgta
     2041 cagaaagtgt gttaaatcca gagaagaaac tggcctactc atgcctctaa aagccccaaa
     2101 agaaattatc ttcttagagg gagaaacact tcccacagaa gtgttaacag aggaagttgt
     2161 cttgaaaact ggtgatttac aaccattaga acaacctact agtgaagctg ttgaagctcc
     2221 attggttggt acaccagttt gtattaacgg gcttatgttg ctcgaaatca aagacacaga
     2281 aaagtactgt gcccttgcac ctaatatgat ggtaacaaac aataccttca cactcaaagg
     2341 cctcgtgccg agaggttctt ggagccatcc gcagtttgaa aaataaacag gccccttttc
     2401 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2461 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2521 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2581 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     2641 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     2701 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     2761 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     2821 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     2881 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     2941 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     3001 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     3061 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     3121 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     3181 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     3241 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3301 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     3361 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     3421 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     3481 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     3541 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     3601 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     3661 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     3721 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3781 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     3841 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3901 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     3961 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     4021 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     4081 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     4141 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     4201 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     4261 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4321 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     4381 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     4441 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     4501 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     4561 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     4621 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     4681 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     4741 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     4801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4861 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     4921 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4981 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5041 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5101 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5161 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5221 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5281 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     5341 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     5401 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     5461 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     5521 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     5581 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     5641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5701 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     5761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5821 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     5881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6061 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     6121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     6181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6301 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     6361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6481 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     6541 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     6601 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     6661 atggtcagtt aga
//
