LOCUS       sequence-291495-        6721 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 6721)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6721)
  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-m4046306
FEATURES             Location/Qualifiers
     source          1..6721
                     /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             2348..2365
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2366..2431
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2476..2494)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2845..3648
                     /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      3731..4713
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4791..4820
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4842..5429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5441..5468)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5492..5683)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5684..6475,6476..6544))
                     /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     6307..6326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6545..6647)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6648..6679)
                     /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 atggcataca cgaggtatgt agacaataac ttttgcggtc ctgatggata
      481 cccactggaa tgtatcaaag atttgttagc tcgtgccggt aaggctagct gtaccctttc
      541 tgagcaattg gacttcatag acactaagag aggtgtttac tgttgtcgcg aacatgaaca
      601 cgaaattgcc tggtataccg agagatcaga aaaatcctac gaattacaga ctccattcga
      661 aattaagttg gctaagaagt ttgatacatt caacggcgaa tgccccaatt tcgtctttcc
      721 attgaactct atcattaaaa ctatccaacc gagagtcgag aagaagaaat tggatggttt
      781 tatgggtaga attagatcgg tgtacccagt tgcttcccca aatgaatgta accaaatgtg
      841 tctatctact ttgatgaagt gtgaccactg tggtgaaacc agttggcaaa caggcgactt
      901 cgttaaggct acatgtgaat tttgcggtac cgagaacctc actaaggaag gtgcgactac
      961 ttgtggttat ttacctcaaa atgcagttgt caagatttac tgtccagctt gccataactc
     1021 tgaagttgga cctgaacaca gcttggctga ataccacaac gaatccggtt taaagaccat
     1081 ccttcgaaaa ggtggtcgta ccatagcctt cgggggttgc gtcttctcat atgttggttg
     1141 tcataataag tgtgcctatt gggttccaag agcttctgct aatatcggat gtaaccacac
     1201 gggtgtggtc ggtgaaggct ctgagggttt gaacgataac ttgttggaaa tacttcagaa
     1261 ggaaaaggtc aacattaaca tcgttggtga ctttaaattg aatgaagaaa ttgctattat
     1321 cctagcgtcc ttctcggcct caacttccgc ttttgtagaa actgttaagg gtcttgatta
     1381 caaagctttc aagcaaatcg tagaatcatg cggtaatttc aaggttacca aggggaaagc
     1441 caaaaaaggt gcctggaaca ttggtgaaca aaagtctata ctgagtccat tatatgcttt
     1501 cgcatctgaa gctgctagag tcgtcagatc tattttttcc agaactctcg aaaccgctca
     1561 aaattcagtg agagttttgc aaaaagcagc cattactatc ttggacggta tttcccagta
     1621 ctctttgagg ttaatcgatg ctatgatgtt tacctccgac ttggctacta ataacctggt
     1681 cgtcatggct tacataaccg gtggcgttgt tcaattgaca tctcaatggc taactaatat
     1741 attcggtact gtttatgaaa agctcaagcc agtcttggat tggttggagg aaaagttcaa
     1801 ggagggcgtt gagtttttac gggatggttg ggaaatcgtt aagttcattt ctacatgtgc
     1861 ctgtgaaatt gtcggtggtc aaatcgtgac ctgtgctaag gaaattaagg agtcggttca
     1921 aactttcttt aagttggtga acaaattttt agctctctgt gctgattcga ttatcatcgg
     1981 aggagcgaaa ttgaaggcac tgaacttggg tgaaaccttc gtcacccact ccaaaggtct
     2041 gtaccgtaaa tgtgttaaat caagagaaga aactggtttg ctaatgcctt tgaaggcccc
     2101 gaaggaaatt atcttcttgg aaggtgaaac attaccaaca gaggttttaa cggaagaggt
     2161 agttttgaag acgggggatt tgcagccgtt agaacaacca acctctgaag ccgtggaagc
     2221 tcccctagtt ggaacgccag tttgcattaa cggtctgatg ttattggaaa tcaaggacac
     2281 cgaaaagtac tgtgctctcg ctccaaacat gatggtcact aacaatacct tcactttgaa
     2341 gggtggtctc gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca
     2401 tgatattgat tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat
     2461 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     2521 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     2581 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     2641 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     2701 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     2761 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     2821 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     2881 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     2941 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     3001 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     3061 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     3121 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     3181 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     3241 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     3301 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     3361 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     3421 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     3481 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     3541 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     3601 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     3661 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     3721 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     3781 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     3841 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     3901 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3961 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     4021 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     4081 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     4141 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     4201 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     4261 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     4321 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     4381 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     4441 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     4501 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     4561 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     4621 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     4681 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     4741 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     4801 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     4861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4921 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     4981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     5341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     5401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     5461 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     5521 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     5581 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     5641 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     5701 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5761 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     5821 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     5881 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5941 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6001 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6061 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6121 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6181 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     6241 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6301 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6361 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6421 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6481 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6541 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     6601 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     6661 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     6721 a
//
