LOCUS       sequence-287632-        4798 bp    DNA     circular SYN 15-APR-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 4798)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4798)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046359
FEATURES             Location/Qualifiers
     source          1..4798
                     /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"
     CDS             491..508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(553..571)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             922..1725
                     /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      1808..2790
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2868..2897
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2919..3506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3518..3545)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3569..3760)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3761..4552,4553..4621))
                     /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     4384..4403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4622..4724)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4725..4756)
                     /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 caccatcatc accatcatta aacaggcccc ttttcctttg tcgatatcat
      541 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
      601 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
      661 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc
      721 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
      781 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
      841 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
      901 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
      961 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     1021 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     1081 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     1141 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     1201 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     1261 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     1321 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     1381 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     1441 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     1501 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     1561 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     1621 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     1681 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     1741 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     1801 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     1861 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     1921 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     1981 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     2041 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     2101 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     2161 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     2221 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     2281 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     2341 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     2401 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     2461 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     2521 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     2581 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     2641 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     2701 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca
     2761 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc
     2821 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac
     2881 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc
     2941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3001 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata
     3061 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3121 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3181 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3241 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3301 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3361 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt
     3421 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct
     3481 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt
     3541 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg
     3601 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     3661 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3721 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg
     3781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3841 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc
     3901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg
     3961 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4201 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4261 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa
     4321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4441 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4621 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     4681 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     4741 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
