LOCUS       sequence-298134-        6172 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 6172)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6172)
  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-m4046558
FEATURES             Location/Qualifiers
     source          1..6172
                     /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             767..784
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1927..1945)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2296..3099
                     /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      3182..4164
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4242..4271
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4293..4880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4892..4919)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4943..5134)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5135..5926,5927..5995))
                     /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     5758..5777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5996..6098)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6099..6130)
                     /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 atgaatgagc ttagtcctgt tgcactacga cagatgtctt gtgctgccgg
      481 tactacacaa actgcttgca ctgatgacaa tgcgttagct tactacaaca caacaaaggg
      541 aggtaggttt gtacttgcac tgttatccga tttacaggat ttgaaatggg ctagattccc
      601 taagagtgat ggaactggta ctatctatac agaactggaa ccaccttgta ggtttgttac
      661 agacacacct aaaggtccta aagtgaagta tttatacttt attaaaggat taaacaacct
      721 aaatagaggt atggtacttg gtagtttagc tgccacagta cgtctactcg tgccgagagg
      781 ttctaaaatc gaggaaggta agctagtcat ttggataaat ggagataagg gttacaacgg
      841 tttggcagaa gtaggcaaga aatttgaaaa ggacacaggt atcaaggtga cggttgaaca
      901 tccagacaag ttagaagaaa aattccctca agttgctgct actggtgatg gtcccgacat
      961 tattttctgg gcccacgatc gttttggtgg atatgctcag tcaggtttgc tggctgagat
     1021 caccccagac aaggccttcc aagataaatt gtacccattc acttgggatg ctgtcagata
     1081 caacggtaag cttatagcat atccaattgc tgttgaagcc ttatcgttga tctacaataa
     1141 ggacttgtta cctaacccac caaagacatg ggaagaaatt ccagctttgg ataaagagct
     1201 aaaggccaag ggaaaaagcg ctttgatgtt taacttacaa gaaccttatt tcacctggcc
     1261 attaatcgct gctgacggtg gttacgcatt taaatacgaa aacggtaagt atgacattaa
     1321 ggatgtcggc gttgataatg ctggtgcaaa ggctggtttg accttcttgg ttgatttaat
     1381 aaagaataaa cacatgaacg ctgacactga ctactctatc gccgaagccg ctttcaacaa
     1441 gggtgaaact gctatgacca taaatggtcc atgggcctgg agtaacattg atacttctaa
     1501 agtaaactat ggtgtcactg ttttgcctac attcaaaggc caaccatcca agccctttgt
     1561 cggtgttcta tccgctggca tcaatgctgc ttctccaaac aaggagctag ctaaagaatt
     1621 tttggaaaat tacttgttaa ctgacgaagg tttggaggcc gttaacaagg acaagccatt
     1681 gggtgctgtc gctttgaaat cttacgaaga agaattggct aaggatccca gaattgccgc
     1741 taccatggag aacgcccaaa agggtgaaat tatgccaaac atcccacaaa tgtcagcctt
     1801 ctggtacgca gttaggactg ccgtgattaa tgctgcttcc ggtagacaaa ccgtcgacga
     1861 agcattaaaa gatgctcaaa cgtaaacagg ccccttttcc tttgtcgata tcatgtaatt
     1921 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
     1981 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
     2041 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca
     2101 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
     2161 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     2221 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     2281 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     2341 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     2401 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     2461 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     2521 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     2581 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     2641 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     2701 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     2761 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     2821 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     2881 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     2941 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     3001 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     3061 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     3121 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     3181 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     3241 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     3301 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     3361 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     3421 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     3481 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     3541 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     3601 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     3661 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     3721 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     3781 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     3841 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     3901 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     3961 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     4021 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     4081 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
     4141 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
     4201 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     4261 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4381 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
     4441 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4501 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4561 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4621 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4681 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4741 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt
     4801 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     4861 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     4921 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     4981 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     5041 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     5101 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac
     5161 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5221 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5281 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca
     5341 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5401 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5461 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5521 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5581 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5641 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt
     5701 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5761 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5821 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5881 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5941 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag
     6001 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     6061 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag
     6121 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
