LOCUS       sequence-298475-        6637 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 6637)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6637)
  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-m4046660
FEATURES             Location/Qualifiers
     source          1..6637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2392..2410)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2761..3564
                     /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      3647..4629
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4707..4736
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4758..5345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5357..5384)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5408..5599)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5600..6391,6392..6460))
                     /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     6223..6242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6461..6563)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6564..6595)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggcataca cgaggtatgt agacaataac ttttgcggtc ctgatggata
      481 cccactggaa tgtatcaaag atttgttagc tcgtgccggt aaggctagct gtaccctttc
      541 tgagcaattg gacttcatag acactaagag aggtgtttac tgttgtcgcg aacatgaaca
      601 cgaaattgcc tggtataccg agagatcaga aaaatcctac gaattacaga ctccattcga
      661 aattaagttg gctaagaagt ttgatacatt caacggcgaa tgccccaatt tcgtctttcc
      721 attgaactct atcattaaaa ctatccaacc gagagtcgag aagaagaaat tggatggttt
      781 tatgggtaga attagatcgg tgtacccagt tgcttcccca aatgaatgta accaaatgtg
      841 tctatctact ttgatgaagt gtgaccactg tggtgaaacc agttggcaaa caggcgactt
      901 cgttaaggct acatgtgaat tttgcggtac cgagaacctc actaaggaag gtgcgactac
      961 ttgtggttat ttacctcaaa atgcagttgt caagatttac tgtccagctt gccataactc
     1021 tgaagttgga cctgaacaca gcttggctga ataccacaac gaatccggtt taaagaccat
     1081 ccttcgaaaa ggtggtcgta ccatagcctt cgggggttgc gtcttctcat atgttggttg
     1141 tcataataag tgtgcctatt gggttccaag agcttctgct aatatcggat gtaaccacac
     1201 gggtgtggtc ggtgaaggct ctgagggttt gaacgataac ttgttggaaa tacttcagaa
     1261 ggaaaaggtc aacattaaca tcgttggtga ctttaaattg aatgaagaaa ttgctattat
     1321 cctagcgtcc ttctcggcct caacttccgc ttttgtagaa actgttaagg gtcttgatta
     1381 caaagctttc aagcaaatcg tagaatcatg cggtaatttc aaggttacca aggggaaagc
     1441 caaaaaaggt gcctggaaca ttggtgaaca aaagtctata ctgagtccat tatatgcttt
     1501 cgcatctgaa gctgctagag tcgtcagatc tattttttcc agaactctcg aaaccgctca
     1561 aaattcagtg agagttttgc aaaaagcagc cattactatc ttggacggta tttcccagta
     1621 ctctttgagg ttaatcgatg ctatgatgtt tacctccgac ttggctacta ataacctggt
     1681 cgtcatggct tacataaccg gtggcgttgt tcaattgaca tctcaatggc taactaatat
     1741 attcggtact gtttatgaaa agctcaagcc agtcttggat tggttggagg aaaagttcaa
     1801 ggagggcgtt gagtttttac gggatggttg ggaaatcgtt aagttcattt ctacatgtgc
     1861 ctgtgaaatt gtcggtggtc aaatcgtgac ctgtgctaag gaaattaagg agtcggttca
     1921 aactttcttt aagttggtga acaaattttt agctctctgt gctgattcga ttatcatcgg
     1981 aggagcgaaa ttgaaggcac tgaacttggg tgaaaccttc gtcacccact ccaaaggtct
     2041 gtaccgtaaa tgtgttaaat caagagaaga aactggtttg ctaatgcctt tgaaggcccc
     2101 gaaggaaatt atcttcttgg aaggtgaaac attaccaaca gaggttttaa cggaagaggt
     2161 agttttgaag acgggggatt tgcagccgtt agaacaacca acctctgaag ccgtggaagc
     2221 tcccctagtt ggaacgccag tttgcattaa cggtctgatg ttattggaaa tcaaggacac
     2281 cgaaaagtac tgtgctctcg ctccaaacat gatggtcact aacaatacct tcactttgaa
     2341 gggtggttaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     2401 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     2461 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     2521 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     2581 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     2641 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     2701 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     2761 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     2821 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     2881 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     2941 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     3001 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     3061 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     3121 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     3181 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     3241 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     3301 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     3361 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     3421 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     3481 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     3541 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     3601 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     3661 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3721 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     3781 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3841 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     3901 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     3961 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     4021 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     4081 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     4141 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     4201 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     4261 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     4321 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     4381 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     4441 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     4501 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     4561 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     4621 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     4681 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     4741 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4801 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4861 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4921 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4981 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5041 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5101 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5161 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5221 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     5281 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     5341 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     5401 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     5461 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     5521 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     5581 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     5641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     5701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5761 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     5821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     6061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6121 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     6301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     6481 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     6541 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     6601 ctatggaggt caggtatgat ttaaatggtc agttaga
//
