LOCUS       sequence-291675-        6631 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 6631)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6631)
  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..6631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2386..2404)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2755..3558
                     /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      3641..4623
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4701..4730
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4752..5339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5351..5378)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5402..5593)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5594..6385,6386..6454))
                     /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     6217..6236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6455..6557)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6558..6589)
                     /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 ctaaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     2401 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     2461 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     2521 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     2581 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     2641 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     2701 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     2761 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     2821 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     2881 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     2941 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     3001 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     3061 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     3121 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     3181 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     3241 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     3301 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     3361 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     3421 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     3481 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     3541 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     3601 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     3661 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     3721 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     3781 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     3841 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     3901 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     3961 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     4021 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     4081 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     4141 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     4201 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     4261 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     4321 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     4381 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     4441 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     4501 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     4561 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     4621 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     4681 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     4741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     4861 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5221 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5281 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5341 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     5401 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     5461 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     5521 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     5581 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     5701 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5761 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     5821 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5881 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5941 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6001 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     6061 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6121 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     6301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6421 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     6481 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     6541 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     6601 aggtcaggta tgatttaaat ggtcagttag a
//
