LOCUS       sequence-297958-        6714 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 6714)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6714)
  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-m4046520
FEATURES             Location/Qualifiers
     source          1..6714
                     /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..2424
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2469..2487)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2838..3641
                     /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      3724..4706
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4784..4813
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4835..5422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5434..5461)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5485..5676)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5677..6468,6469..6537))
                     /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     6300..6319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6538..6640)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6641..6672)
                     /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 ggttctcacc atcatcacca tcattaaaca ggcccctttt cctttgtcga tatcatgtaa
     2461 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     2521 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     2581 naagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     2641 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     2701 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     2761 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     2821 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     2881 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     2941 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     3001 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     3061 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     3121 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     3181 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     3241 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     3301 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     3361 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     3421 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     3481 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     3541 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     3601 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     3661 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     3721 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     3781 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     3841 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     3901 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     3961 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     4021 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     4081 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     4141 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     4201 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     4261 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     4321 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     4381 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     4441 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     4501 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     4561 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     4621 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     4681 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     4741 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     4801 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4861 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4921 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4981 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5041 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5101 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5161 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5221 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5281 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     5341 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     5401 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     5461 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     5521 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5581 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5641 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     5701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5761 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     5881 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6121 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6181 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     6241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6361 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     6541 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     6601 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     6661 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
