LOCUS       sequence-291571-        6762 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 6762)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6762)
  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-m4046369
FEATURES             Location/Qualifiers
     source          1..6762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2389..2406
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2407..2472
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2517..2535)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2886..3689
                     /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      3772..4754
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4832..4861
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4883..5470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5482..5509)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5533..5724)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5725..6516,6517..6585))
                     /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     6348..6367
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6586..6688)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6689..6720)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 tacactcgct atgtcgataa caacttctgt ggccctgatg gctaccctct tgagtgcatt
      541 aaagaccttc tagcacgtgc tggtaaagct tcatgcactt tgtccgaaca actggacttt
      601 attgacacta agaggggtgt atactgctgc cgtgaacatg agcatgaaat tgcttggtac
      661 acggaacgtt ctgaaaagag ctatgaattg cagacacctt ttgaaattaa attggcaaag
      721 aaatttgaca ccttcaatgg ggaatgtcca aattttgtat ttcccttaaa ttccataatc
      781 aagactattc aaccaagggt tgaaaagaaa aagcttgatg gctttatggg tagaattcga
      841 tctgtctatc cagttgcgtc accaaatgaa tgcaaccaaa tgtgcctttc aactctcatg
      901 aagtgtgatc attgtggtga aacttcatgg cagacgggcg attttgttaa agccacttgc
      961 gaattttgtg gcactgagaa tttgactaaa gaaggtgcca ctacttgtgg ttacttaccc
     1021 caaaatgctg ttgttaaaat ttattgtcca gcatgtcaca attcagaagt aggacctgag
     1081 catagtcttg ccgaatacca taatgaatct ggcttgaaaa ccattcttcg taagggtggt
     1141 cgcactattg cctttggagg ctgtgtgttc tcttatgttg gttgccataa caagtgtgcc
     1201 tattgggttc cacgtgctag cgctaacata ggttgtaacc atacaggtgt tgttggagaa
     1261 ggttccgaag gtcttaatga caaccttctt gaaatactcc aaaaagagaa agtcaacatc
     1321 aatattgttg gtgactttaa acttaatgaa gagatcgcca ttattttggc atctttttct
     1381 gcttccacaa gtgcttttgt ggaaactgtg aaaggtttgg attataaagc attcaaacaa
     1441 attgttgaat cctgtggtaa ttttaaagtt acaaaaggaa aagctaaaaa aggtgcctgg
     1501 aatattggtg aacagaaatc aatactgagt cctctttatg catttgcatc agaggctgct
     1561 cgtgttgtac gatcaatttt ctcccgcact cttgaaactg ctcaaaattc tgtgcgtgtt
     1621 ttacagaagg ccgctataac aatactagat ggaatttcac agtattcact gagactcatt
     1681 gatgctatga tgttcacatc tgatttggct actaacaatc tagttgtaat ggcctacatt
     1741 acaggtggtg ttgttcagtt gacttcgcag tggctaacta acatctttgg cactgtttat
     1801 gaaaaactca aacccgtcct tgattggctt gaagagaagt ttaaggaagg tgtagagttt
     1861 cttagagacg gttgggaaat tgttaaattt atctcaacct gtgcttgtga aattgtcggt
     1921 ggacaaattg tcacctgtgc aaaggaaatt aaggagagtg ttcagacatt ctttaagctt
     1981 gtaaataaat ttttggcttt gtgtgctgac tctatcatta ttggtggagc taaacttaaa
     2041 gccttgaatt taggtgaaac atttgtcacg cactcaaagg gattgtacag aaagtgtgtt
     2101 aaatccagag aagaaactgg cctactcatg cctctaaaag ccccaaaaga aattatcttc
     2161 ttagagggag aaacacttcc cacagaagtg ttaacagagg aagttgtctt gaaaactggt
     2221 gatttacaac cattagaaca acctactagt gaagctgttg aagctccatt ggttggtaca
     2281 ccagtttgta ttaacgggct tatgttgctc gaaatcaaag acacagaaaa gtactgtgcc
     2341 cttgcaccta atatgatggt aacaaacaat accttcacac tcaaaggcct cgtgccgaga
     2401 ggttctgatt ataaagatca tgatggtgat tataaagatc atgatattga ttataaagat
     2461 gatgatgata aataaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     2521 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     2581 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     2641 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     2701 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     2761 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     2821 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     2881 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     2941 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     3001 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     3061 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     3121 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     3181 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     3241 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     3301 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     3361 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     3421 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     3481 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     3541 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     3601 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     3661 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     3721 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     3781 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3841 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     3901 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     3961 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     4021 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     4081 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     4141 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     4201 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     4261 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     4321 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     4381 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     4441 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     4501 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     4561 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     4621 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     4681 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     4741 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     4801 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     4861 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4921 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4981 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5041 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5101 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5161 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5221 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5281 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5341 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     5401 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     5461 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     5521 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     5581 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     5641 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     5701 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     5761 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5821 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5881 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     5941 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6001 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6061 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6121 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     6181 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6241 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6301 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6361 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     6421 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6481 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6541 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     6601 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     6661 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     6721 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
