LOCUS       sequence-298963-        6720 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 6720)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6720)
  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-m4046441
FEATURES             Location/Qualifiers
     source          1..6720
                     /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..2430
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2475..2493)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2844..3647
                     /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      3730..4712
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4790..4819
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4841..5428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5440..5467)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5491..5682)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5683..6474,6475..6543))
                     /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     6306..6325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6544..6646)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6647..6678)
                     /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 ctcaccatca tcaccatcat taaacaggcc ccttttcctt tgtcgatatc
     2461 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     2521 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     2581 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     2641 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     2701 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     2761 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     2821 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     2881 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     2941 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     3001 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     3061 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     3121 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     3181 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     3241 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     3301 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     3361 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     3421 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     3481 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     3541 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     3601 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     3661 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     3721 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     3781 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     3841 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     3901 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     3961 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     4021 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     4081 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     4141 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     4201 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     4261 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     4321 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     4381 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     4441 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     4501 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     4561 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     4621 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     4681 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     4741 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     4801 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     4861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4921 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     4981 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5041 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5101 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5161 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5221 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5281 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     5341 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     5401 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     5461 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     5521 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     5581 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     5641 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     5701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5761 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     5821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     5881 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6121 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6181 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     6241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6361 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6541 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     6601 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     6661 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
