LOCUS       sequence-287591-        5671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5671)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5671)
  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-m4046360
FEATURES             Location/Qualifiers
     source          1..5671
                     /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             1298..1315
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1316..1381
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1426..1444)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1795..2598
                     /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      2681..3663
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3741..3770
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3792..4379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4391..4418)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4442..4633)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4634..5425,5426..5494))
                     /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     5257..5276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5495..5597)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5598..5629)
                     /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 atggcagtga aaagaacaat caagggtaca caccactggt tgttactcac
      481 aattttgact tcacttttag ttttagtcca gagtactcaa tggtctttgt tctttttttt
      541 gtatgaaaat gcctttttac cttttgctat gggtattatt gctatgtctg cttttgcaat
      601 gatgtttgtc aaacataagc atgcatttct ctgtttgttt ttgttacctt ctcttgccac
      661 tgtagcttat tttaatatgg tctatatgcc tgctagttgg gtgatgcgta ttatgacatg
      721 gttggatatg gttgatacta gtttgtctgg ttttaagcta aaagactgtg ttatgtatgc
      781 atcagctgta gtgttactaa tccttatgac agcaagaact gtgtatgatg atggtgctag
      841 gagagtgtgg acacttatga atgtcttgac actcgtttat aaagtttatt atggtaatgc
      901 tttagatcaa gccatttcca tgtgggctct tataatctct gttacttcta actactcagg
      961 tgtagttaca actgtcatgt ttttggccag aggtattgtt tttatgtgtg ttgagtattg
     1021 ccctattttc ttcataactg gtaatacact tcagtgtata atgctagttt attgtttctt
     1081 aggctatttt tgtacttgtt actttggcct cttttgttta ctcaaccgct actttagact
     1141 gactcttggt gtttatgatt acttagtttc tacacaggag tttagatata tgaattcaca
     1201 gggactactc ccacccaaga atagcataga tgccttcaaa ctcaacatta aattgttggg
     1261 tgttggtggc aaaccttgta tcaaagtagc cactgtactc gtgccgagag gttctgatta
     1321 taaagatcat gatggtgatt ataaagatca tgatattgat tataaagatg atgatgataa
     1381 ataaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     1441 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1501 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     1561 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     1621 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     1681 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     1741 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     1801 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     1861 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     1921 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     1981 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     2041 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     2101 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     2161 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     2221 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     2281 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     2341 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     2401 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     2461 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     2521 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     2581 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     2641 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     2701 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2761 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2821 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2881 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     2941 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     3001 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     3061 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     3121 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     3181 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     3241 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     3301 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     3361 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     3421 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     3481 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     3541 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     3601 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     3661 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     3721 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     3781 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3841 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     3901 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3961 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4021 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4081 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4141 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4201 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4261 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4321 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4381 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     4441 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4501 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4561 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4621 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4681 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     4741 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4801 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     4861 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4921 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4981 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5041 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     5101 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5161 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5221 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5281 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     5341 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5401 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5461 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     5521 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     5581 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     5641 aggtcaggta tgatttaaat ggtcagttag a
//
