LOCUS       sequence-297825-        6679 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 6679)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6679)
  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-m4046368
FEATURES             Location/Qualifiers
     source          1..6679
                     /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..2389
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2434..2452)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2803..3606
                     /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      3689..4671
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4749..4778
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4800..5387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5399..5426)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5450..5641)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5642..6433,6434..6502))
                     /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     6265..6284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6503..6605)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6606..6637)
                     /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 gttcttggag ccatccgcag tttgaaaaat aaacaggccc
     2401 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     2461 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     2521 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     2581 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     2641 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     2701 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     2761 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     2821 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     2881 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     2941 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     3001 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     3061 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     3121 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     3181 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     3241 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     3301 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     3361 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     3421 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     3481 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     3541 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     3601 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     3661 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     3721 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     3781 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     3841 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3901 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     3961 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     4021 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     4081 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     4141 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     4201 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     4261 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     4321 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     4381 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     4441 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     4501 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     4561 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     4621 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     4681 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     4741 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     4801 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4861 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     4921 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5161 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5281 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5341 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5401 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     5461 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     5521 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     5581 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     5641 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5701 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     5761 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5821 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     5881 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5941 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6001 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6061 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     6121 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     6181 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6241 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6301 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     6361 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6421 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6481 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     6541 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     6601 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     6661 atttaaatgg tcagttaga
//
