LOCUS       sequence-299995-        6684 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 6684)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6684)
  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-m4046672
FEATURES             Location/Qualifiers
     source          1..6684
                     /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"
     primer_bind     complement(2439..2457)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2808..3611
                     /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      3694..4676
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4754..4783
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4805..5392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5404..5431)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5455..5646)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5647..6438,6439..6507))
                     /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     6270..6289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6508..6610)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6611..6642)
                     /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 tggttaaaca
     2401 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     2461 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     2521 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     2581 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     2641 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     2701 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     2761 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     2821 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     2881 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     2941 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     3001 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     3061 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     3121 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     3181 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     3241 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     3301 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     3361 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     3421 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     3481 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     3541 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     3601 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     3661 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     3721 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3781 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     3841 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     3901 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     3961 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     4021 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     4081 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     4141 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     4201 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     4261 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     4321 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     4381 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     4441 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     4501 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     4561 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     4621 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     4681 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     4741 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     4801 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4861 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     4921 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5161 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5281 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5341 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5401 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     5461 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     5521 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     5581 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5641 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5701 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     5761 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5821 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     5881 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5941 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6001 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6061 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     6121 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     6181 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6241 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6301 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     6361 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6421 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6481 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     6541 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     6601 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     6661 gtatgattta aatggtcagt taga
//
