LOCUS       sequence-287662-        6768 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 6768)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6768)
  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-m4046549
FEATURES             Location/Qualifiers
     source          1..6768
                     /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             2395..2412
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2413..2478
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2523..2541)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2892..3695
                     /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      3778..4760
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4838..4867
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4889..5476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5488..5515)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5539..5730)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5731..6522,6523..6591))
                     /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     6354..6373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6592..6694)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6695..6726)
                     /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 gcatacacga ggtatgtaga caataacttt tgcggtcctg atggataccc actggaatgt
      541 atcaaagatt tgttagctcg tgccggtaag gctagctgta ccctttctga gcaattggac
      601 ttcatagaca ctaagagagg tgtttactgt tgtcgcgaac atgaacacga aattgcctgg
      661 tataccgaga gatcagaaaa atcctacgaa ttacagactc cattcgaaat taagttggct
      721 aagaagtttg atacattcaa cggcgaatgc cccaatttcg tctttccatt gaactctatc
      781 attaaaacta tccaaccgag agtcgagaag aagaaattgg atggttttat gggtagaatt
      841 agatcggtgt acccagttgc ttccccaaat gaatgtaacc aaatgtgtct atctactttg
      901 atgaagtgtg accactgtgg tgaaaccagt tggcaaacag gcgacttcgt taaggctaca
      961 tgtgaatttt gcggtaccga gaacctcact aaggaaggtg cgactacttg tggttattta
     1021 cctcaaaatg cagttgtcaa gatttactgt ccagcttgcc ataactctga agttggacct
     1081 gaacacagct tggctgaata ccacaacgaa tccggtttaa agaccatcct tcgaaaaggt
     1141 ggtcgtacca tagccttcgg gggttgcgtc ttctcatatg ttggttgtca taataagtgt
     1201 gcctattggg ttccaagagc ttctgctaat atcggatgta accacacggg tgtggtcggt
     1261 gaaggctctg agggtttgaa cgataacttg ttggaaatac ttcagaagga aaaggtcaac
     1321 attaacatcg ttggtgactt taaattgaat gaagaaattg ctattatcct agcgtccttc
     1381 tcggcctcaa cttccgcttt tgtagaaact gttaagggtc ttgattacaa agctttcaag
     1441 caaatcgtag aatcatgcgg taatttcaag gttaccaagg ggaaagccaa aaaaggtgcc
     1501 tggaacattg gtgaacaaaa gtctatactg agtccattat atgctttcgc atctgaagct
     1561 gctagagtcg tcagatctat tttttccaga actctcgaaa ccgctcaaaa ttcagtgaga
     1621 gttttgcaaa aagcagccat tactatcttg gacggtattt cccagtactc tttgaggtta
     1681 atcgatgcta tgatgtttac ctccgacttg gctactaata acctggtcgt catggcttac
     1741 ataaccggtg gcgttgttca attgacatct caatggctaa ctaatatatt cggtactgtt
     1801 tatgaaaagc tcaagccagt cttggattgg ttggaggaaa agttcaagga gggcgttgag
     1861 tttttacggg atggttggga aatcgttaag ttcatttcta catgtgcctg tgaaattgtc
     1921 ggtggtcaaa tcgtgacctg tgctaaggaa attaaggagt cggttcaaac tttctttaag
     1981 ttggtgaaca aatttttagc tctctgtgct gattcgatta tcatcggagg agcgaaattg
     2041 aaggcactga acttgggtga aaccttcgtc acccactcca aaggtctgta ccgtaaatgt
     2101 gttaaatcaa gagaagaaac tggtttgcta atgcctttga aggccccgaa ggaaattatc
     2161 ttcttggaag gtgaaacatt accaacagag gttttaacgg aagaggtagt tttgaagacg
     2221 ggggatttgc agccgttaga acaaccaacc tctgaagccg tggaagctcc cctagttgga
     2281 acgccagttt gcattaacgg tctgatgtta ttggaaatca aggacaccga aaagtactgt
     2341 gctctcgctc caaacatgat ggtcactaac aataccttca ctttgaaggg tggtctcgtg
     2401 ccgagaggtt ctgattataa agatcatgat ggtgattata aagatcatga tattgattat
     2461 aaagatgatg atgataaata aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     2521 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     2581 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     2641 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     2701 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     2761 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     2821 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     2881 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     2941 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     3001 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     3061 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     3121 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     3181 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     3241 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     3301 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     3361 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     3421 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     3481 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     3541 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     3601 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     3661 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     3721 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     3781 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     3841 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     3901 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     3961 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     4021 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     4081 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     4141 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     4201 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     4261 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     4321 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     4381 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     4441 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     4501 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     4561 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     4621 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     4681 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     4741 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     4801 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     4861 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4921 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4981 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5041 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5101 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5161 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5221 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5281 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5341 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     5401 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     5461 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     5521 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     5581 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     5641 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     5701 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     5761 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5821 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5881 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     5941 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6001 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6061 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6121 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     6181 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6241 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6301 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6361 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     6421 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6481 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6541 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     6601 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     6661 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     6721 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
