LOCUS       sequence-298103-        7747 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 7747)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7747)
  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-m4046164
FEATURES             Location/Qualifiers
     source          1..7747
                     /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"
     primer_bind     complement(3502..3520)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3871..4674
                     /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      4757..5739
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5817..5846
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5868..6455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6467..6494)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6518..6709)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6710..7501,7502..7570))
                     /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     7333..7352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7571..7673)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7674..7705)
                     /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 agaggttcta aaatcgagga aggtaagcta gtcatttgga taaatggaga
     2401 taagggttac aacggtttgg cagaagtagg caagaaattt gaaaaggaca caggtatcaa
     2461 ggtgacggtt gaacatccag acaagttaga agaaaaattc cctcaagttg ctgctactgg
     2521 tgatggtccc gacattattt tctgggccca cgatcgtttt ggtggatatg ctcagtcagg
     2581 tttgctggct gagatcaccc cagacaaggc cttccaagat aaattgtacc cattcacttg
     2641 ggatgctgtc agatacaacg gtaagcttat agcatatcca attgctgttg aagccttatc
     2701 gttgatctac aataaggact tgttacctaa cccaccaaag acatgggaag aaattccagc
     2761 tttggataaa gagctaaagg ccaagggaaa aagcgctttg atgtttaact tacaagaacc
     2821 ttatttcacc tggccattaa tcgctgctga cggtggttac gcatttaaat acgaaaacgg
     2881 taagtatgac attaaggatg tcggcgttga taatgctggt gcaaaggctg gtttgacctt
     2941 cttggttgat ttaataaaga ataaacacat gaacgctgac actgactact ctatcgccga
     3001 agccgctttc aacaagggtg aaactgctat gaccataaat ggtccatggg cctggagtaa
     3061 cattgatact tctaaagtaa actatggtgt cactgttttg cctacattca aaggccaacc
     3121 atccaagccc tttgtcggtg ttctatccgc tggcatcaat gctgcttctc caaacaagga
     3181 gctagctaaa gaatttttgg aaaattactt gttaactgac gaaggtttgg aggccgttaa
     3241 caaggacaag ccattgggtg ctgtcgcttt gaaatcttac gaagaagaat tggctaagga
     3301 tcccagaatt gccgctacca tggagaacgc ccaaaagggt gaaattatgc caaacatccc
     3361 acaaatgtca gccttctggt acgcagttag gactgccgtg attaatgctg cttccggtag
     3421 acaaaccgtc gacgaagcat taaaagatgc tcaaacgtaa acaggcccct tttcctttgt
     3481 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     3541 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     3601 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     3661 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     3721 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     3781 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     3841 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     3901 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     3961 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     4021 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     4081 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     4141 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     4201 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     4261 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     4321 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     4381 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     4441 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     4501 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     4561 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     4621 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     4681 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     4741 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     4801 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     4861 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     4921 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     4981 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     5041 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     5101 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     5161 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     5221 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     5281 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     5341 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     5401 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     5461 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     5521 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     5581 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     5641 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     5701 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     5761 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     5821 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     5881 tttctncgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5941 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     6001 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6061 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     6361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     6421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     6481 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     6541 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     6601 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     6661 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     6721 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6781 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     6841 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6901 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     6961 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7021 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7081 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7141 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7201 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     7261 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7321 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7381 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7441 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7501 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7561 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     7621 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     7681 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     7741 agttaga
//
