LOCUS       sequence-297239-        6673 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 6673)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6673)
  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-m4046450
FEATURES             Location/Qualifiers
     source          1..6673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2348..2365
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2366..2383
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2428..2446)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2797..3600
                     /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      3683..4665
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4743..4772
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4794..5381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5393..5420)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5444..5635)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5636..6427,6428..6496))
                     /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     6259..6278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6497..6599)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6600..6631)
                     /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 gggtggtctc gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc
     2401 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2461 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2521 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2581 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     2641 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     2701 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     2761 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     2821 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     2881 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     2941 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     3001 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     3061 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     3121 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     3181 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     3241 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3301 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     3361 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     3421 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     3481 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     3541 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     3601 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     3661 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     3721 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3781 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     3841 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3901 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     3961 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     4021 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     4081 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     4141 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     4201 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     4261 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4321 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     4381 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     4441 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     4501 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     4561 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     4621 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     4681 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     4741 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     4801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4861 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     4921 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4981 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5041 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5101 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5161 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5221 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5281 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     5341 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     5401 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     5461 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     5521 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     5581 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     5641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5701 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     5761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5821 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     5881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6061 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     6121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     6181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6301 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     6361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6481 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     6541 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     6601 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     6661 atggtcagtt aga
//
