LOCUS       sequence-287622-        5146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5146)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5146)
  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-m4046473
FEATURES             Location/Qualifiers
     source          1..5146
                     /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             773..790
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             791..856
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(901..919)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1270..2073
                     /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      2156..3138
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3216..3245
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3267..3854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3866..3893)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3917..4108)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4109..4900,4901..4969))
                     /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     4732..4751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4970..5072)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5073..5104)
                     /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 atgaacaatg agctttctcc tgttgcttta cggcagatgt cctgcgcagc
      481 tggaacaacg caaactgcct gtactgacga taacgctcta gcttactata acaccactaa
      541 aggtggtaga ttcgtcttgg ccttgttgtc agacctgcaa gatttgaagt gggcgagatt
      601 tccaaagagc gatggcactg gtaccatata caccgaatta gaaccaccct gtcgtttcgt
      661 gaccgacact ccaaaaggtc cgaaggttaa gtatttgtac tttattaagg gtttaaataa
      721 cttgaatcga ggtatggttc tggggtcttt ggctgctaca gtcagactcc aactcgtgcc
      781 gagaggttct gattataaag atcatgatgg tgattataaa gatcatgata ttgattataa
      841 agatgatgat gataaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat
      901 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag
      961 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg
     1021 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg
     1081 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     1141 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     1201 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga
     1261 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     1321 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     1381 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     1441 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     1501 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt
     1561 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     1621 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     1681 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     1741 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     1801 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     1861 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat
     1921 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta
     1981 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     2041 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     2101 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga
     2161 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2221 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     2281 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     2341 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     2401 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     2461 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     2521 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     2581 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     2641 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     2701 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     2761 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     2821 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     2881 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     2941 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     3001 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     3061 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga
     3121 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct
     3181 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt
     3241 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3301 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3361 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3421 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3481 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3541 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3601 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3661 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3721 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc
     3781 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc
     3841 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat
     3901 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3961 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4021 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4081 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct
     4141 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4201 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4261 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg
     4321 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4381 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4441 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4501 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca
     4561 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4621 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4681 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4741 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac
     4801 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4861 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4921 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct
     4981 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     5041 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt
     5101 gactttctgc tatggaggtc aggtatgatt taaatggtca gttaga
//
