LOCUS       sequence-291577-        6715 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 6715)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6715)
  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..6715
                     /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..2425
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2470..2488)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2839..3642
                     /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      3725..4707
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4785..4814
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4836..5423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5435..5462)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5486..5677)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5678..6469,6470..6538))
                     /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     6301..6320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6539..6641)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6642..6673)
                     /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 agaggttctg attataaaga tcatgatggt gattataaag atcatgatat
     2401 tgattataaa gatgatgatg ataaataaac aggccccttt tcctttgtcg atatcatgta
     2461 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     2521 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     2581 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     2641 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     2701 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     2761 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     2821 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     2881 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     2941 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     3001 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     3061 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     3121 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     3181 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     3241 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     3301 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     3361 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     3421 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     3481 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     3541 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     3601 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     3661 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     3721 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     3781 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     3841 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     3901 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     3961 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     4021 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     4081 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     4141 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     4201 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     4261 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     4321 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     4381 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     4441 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     4501 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     4561 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     4621 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     4681 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
     4741 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     4801 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4921 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     4981 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5041 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5101 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5161 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5221 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5281 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc
     5341 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     5401 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     5461 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     5521 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     5581 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     5641 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     5701 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5761 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5821 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     5881 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5941 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6001 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6061 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6121 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6181 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     6241 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6301 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6361 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6421 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6481 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     6541 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     6601 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     6661 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
