LOCUS       sequence-291558-        5092 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 5092)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5092)
  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-m4046338
FEATURES             Location/Qualifiers
     source          1..5092
                     /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"
     CDS             785..802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(847..865)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1216..2019
                     /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      2102..3084
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3162..3191
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3213..3800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3812..3839)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3863..4054)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4055..4846,4847..4915))
                     /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     4678..4697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4916..5018)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5019..5050)
                     /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 ttctcaccat catcaccatc attaaacagg ccccttttcc tttgtcgata tcatgtaatt
      841 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
      901 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
      961 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca
     1021 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
     1081 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     1141 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     1201 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     1261 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     1321 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     1381 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     1441 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     1501 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     1561 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     1621 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     1681 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     1741 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     1801 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     1861 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     1921 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     1981 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     2041 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     2101 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     2161 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     2221 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     2281 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     2341 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     2401 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     2461 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     2521 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     2581 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     2641 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     2701 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     2761 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     2821 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     2881 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     2941 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     3001 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
     3061 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
     3121 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     3181 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3241 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3301 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
     3361 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3421 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3481 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3541 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3601 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3661 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt
     3721 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     3781 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     3841 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3901 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3961 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     4021 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac
     4081 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4141 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4201 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca
     4261 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4321 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4381 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4441 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4501 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4561 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt
     4621 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4681 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4741 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4801 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4861 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag
     4921 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     4981 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag
     5041 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
