LOCUS       sequence-287644-        4792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4792)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4792)
  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-m4046260
FEATURES             Location/Qualifiers
     source          1..4792
                     /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             467..484
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             485..502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(547..565)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             916..1719
                     /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      1802..2784
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2862..2891
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2913..3500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3512..3539)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3563..3754)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3755..4546,4547..4615))
                     /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     4378..4397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4616..4718)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4719..4750)
                     /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 atggctgatg cacaatcgtt tttaaacggg tttgcgctcg tgccgagagg
      481 ttctcaccat catcaccatc attaaacagg ccccttttcc tttgtcgata tcatgtaatt
      541 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
      601 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
      661 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca
      721 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
      781 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
      841 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
      901 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
      961 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     1021 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     1081 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     1141 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     1201 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     1261 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     1321 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     1381 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     1441 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     1501 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     1561 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     1621 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     1681 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     1741 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     1801 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     1861 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     1921 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     1981 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     2041 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     2101 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     2161 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     2221 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     2281 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     2341 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     2401 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     2461 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     2521 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     2581 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     2641 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     2701 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
     2761 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
     2821 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     2881 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3001 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
     3061 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3121 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3181 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3241 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3301 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3361 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt
     3421 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     3481 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     3541 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3601 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3661 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3721 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac
     3781 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3841 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3901 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca
     3961 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4021 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4081 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4141 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4201 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4261 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt
     4321 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4381 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4441 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4501 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4561 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag
     4621 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     4681 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag
     4741 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
