LOCUS       sequence-297592-        7800 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 7800)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7800)
  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-m4046172
FEATURES             Location/Qualifiers
     source          1..7800
                     /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"
     primer_bind     complement(3555..3573)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3924..4727
                     /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      4810..5792
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5870..5899
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5921..6508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6520..6547)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6571..6762)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6763..7554,7555..7623))
                     /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     7386..7405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7624..7726)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7727..7758)
                     /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 ctaaaatcga ggaaggtaag ctagtcattt ggataaatgg agataagggt
     2461 tacaacggtt tggcagaagt aggcaagaaa tttgaaaagg acacaggtat caaggtgacg
     2521 gttgaacatc cagacaagtt agaagaaaaa ttccctcaag ttgctgctac tggtgatggt
     2581 cccgacatta ttttctgggc ccacgatcgt tttggtggat atgctcagtc aggtttgctg
     2641 gctgagatca ccccagacaa ggccttccaa gataaattgt acccattcac ttgggatgct
     2701 gtcagataca acggtaagct tatagcatat ccaattgctg ttgaagcctt atcgttgatc
     2761 tacaataagg acttgttacc taacccacca aagacatggg aagaaattcc agctttggat
     2821 aaagagctaa aggccaaggg aaaaagcgct ttgatgttta acttacaaga accttatttc
     2881 acctggccat taatcgctgc tgacggtggt tacgcattta aatacgaaaa cggtaagtat
     2941 gacattaagg atgtcggcgt tgataatgct ggtgcaaagg ctggtttgac cttcttggtt
     3001 gatttaataa agaataaaca catgaacgct gacactgact actctatcgc cgaagccgct
     3061 ttcaacaagg gtgaaactgc tatgaccata aatggtccat gggcctggag taacattgat
     3121 acttctaaag taaactatgg tgtcactgtt ttgcctacat tcaaaggcca accatccaag
     3181 ccctttgtcg gtgttctatc cgctggcatc aatgctgctt ctccaaacaa ggagctagct
     3241 aaagaatttt tggaaaatta cttgttaact gacgaaggtt tggaggccgt taacaaggac
     3301 aagccattgg gtgctgtcgc tttgaaatct tacgaagaag aattggctaa ggatcccaga
     3361 attgccgcta ccatggagaa cgcccaaaag ggtgaaatta tgccaaacat cccacaaatg
     3421 tcagccttct ggtacgcagt taggactgcc gtgattaatg ctgcttccgg tagacaaacc
     3481 gtcgacgaag cattaaaaga tgctcaaacg taaacaggcc ccttttcctt tgtcgatatc
     3541 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     3601 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     3661 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     3721 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     3781 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     3841 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     3901 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     3961 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     4021 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     4081 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     4141 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     4201 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     4261 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     4321 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     4381 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     4441 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     4501 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     4561 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     4621 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     4681 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     4741 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     4801 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     4861 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     4921 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     4981 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     5041 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     5101 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     5161 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     5221 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     5281 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     5341 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     5401 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     5461 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     5521 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     5581 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     5641 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     5701 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     5761 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     5821 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     5881 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     5941 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6001 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     6061 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6121 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6181 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6241 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6301 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6361 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     6421 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     6481 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     6541 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     6601 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     6661 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     6721 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     6781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6841 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     6901 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     6961 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7021 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7081 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7141 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7201 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7261 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     7321 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7381 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7441 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7501 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7561 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7621 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     7681 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     7741 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
