LOCUS       sequence-298249-        6678 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 6678)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6678)
  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-m4046699
FEATURES             Location/Qualifiers
     source          1..6678
                     /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(2433..2451)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2802..3605
                     /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      3688..4670
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4748..4777
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4799..5386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5398..5425)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5449..5640)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5641..6432,6433..6501))
                     /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     6264..6283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6502..6604)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6605..6636)
                     /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 tcaaaggcta aacaggcccc
     2401 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     2461 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     2521 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     2581 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     2641 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     2701 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     2761 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     2821 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     2881 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     2941 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     3001 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     3061 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     3121 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     3181 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     3241 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     3301 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     3361 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     3421 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     3481 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     3541 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     3601 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     3661 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     3721 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3781 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     3841 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     3901 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     3961 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     4021 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     4081 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     4141 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     4201 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     4261 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     4321 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     4381 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     4441 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     4501 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     4561 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     4621 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     4681 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     4741 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     4801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4861 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     4921 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4981 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5041 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5101 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5161 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5221 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5281 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5341 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5401 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     5461 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     5521 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     5581 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5701 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5821 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6061 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6301 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6481 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     6541 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     6601 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     6661 tttaaatggt cagttaga
//
