LOCUS       sequence-287586-        5878 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 5878)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5878)
  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-m4046321
FEATURES             Location/Qualifiers
     source          1..5878
                     /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             473..490
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1633..1651)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2002..2805
                     /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      2888..3870
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3948..3977
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3999..4586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4598..4625)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4649..4840)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4841..5632,5633..5701))
                     /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     5464..5483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5702..5804)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5805..5836)
                     /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 atgtccgctg acgcacaatc attcctaaat gggtttgctg tactcgtgcc
      481 gagaggttct aaaatcgagg aaggtaagct agtcatttgg ataaatggag ataagggtta
      541 caacggtttg gcagaagtag gcaagaaatt tgaaaaggac acaggtatca aggtgacggt
      601 tgaacatcca gacaagttag aagaaaaatt ccctcaagtt gctgctactg gtgatggtcc
      661 cgacattatt ttctgggccc acgatcgttt tggtggatat gctcagtcag gtttgctggc
      721 tgagatcacc ccagacaagg ccttccaaga taaattgtac ccattcactt gggatgctgt
      781 cagatacaac ggtaagctta tagcatatcc aattgctgtt gaagccttat cgttgatcta
      841 caataaggac ttgttaccta acccaccaaa gacatgggaa gaaattccag ctttggataa
      901 agagctaaag gccaagggaa aaagcgcttt gatgtttaac ttacaagaac cttatttcac
      961 ctggccatta atcgctgctg acggtggtta cgcatttaaa tacgaaaacg gtaagtatga
     1021 cattaaggat gtcggcgttg ataatgctgg tgcaaaggct ggtttgacct tcttggttga
     1081 tttaataaag aataaacaca tgaacgctga cactgactac tctatcgccg aagccgcttt
     1141 caacaagggt gaaactgcta tgaccataaa tggtccatgg gcctggagta acattgatac
     1201 ttctaaagta aactatggtg tcactgtttt gcctacattc aaaggccaac catccaagcc
     1261 ctttgtcggt gttctatccg ctggcatcaa tgctgcttct ccaaacaagg agctagctaa
     1321 agaatttttg gaaaattact tgttaactga cgaaggtttg gaggccgtta acaaggacaa
     1381 gccattgggt gctgtcgctt tgaaatctta cgaagaagaa ttggctaagg atcccagaat
     1441 tgccgctacc atggagaacg cccaaaaggg tgaaattatg ccaaacatcc cacaaatgtc
     1501 agccttctgg tacgcagtta ggactgccgt gattaatgct gcttccggta gacaaaccgt
     1561 cgacgaagca ttaaaagatg ctcaaacgta aacaggcccc ttttcctttg tcgatatcat
     1621 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
     1681 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
     1741 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc
     1801 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
     1861 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     1921 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
     1981 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     2041 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     2101 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     2161 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     2221 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     2281 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     2341 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     2401 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     2461 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     2521 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     2581 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     2641 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     2701 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     2761 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     2821 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     2881 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     2941 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     3001 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     3061 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     3121 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     3181 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     3241 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     3301 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     3361 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     3421 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     3481 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     3541 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     3601 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     3661 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     3721 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     3781 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca
     3841 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc
     3901 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac
     3961 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc
     4021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4081 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata
     4141 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4201 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4261 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4321 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4381 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4441 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt
     4501 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct
     4561 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt
     4621 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg
     4681 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     4741 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4801 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg
     4861 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4921 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc
     4981 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg
     5041 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5101 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5161 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5221 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5281 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5341 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa
     5401 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5461 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5521 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5581 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5641 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5701 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     5761 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     5821 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
