LOCUS       sequence-300492-        6703 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 6703)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6703)
  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-m4046274
FEATURES             Location/Qualifiers
     source          1..6703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      211..1193
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      1271..1300
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1322..1909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1921..1948)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1972..2163)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(2164..2955,2956..3024))
                     /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     2787..2806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3025..3127)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3128..3159)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3205..3618
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4499..4516
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(5659..5677)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             join(6028..6703,1..128)
                     /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"
ORIGIN
        1 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
       61 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
      121 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
      181 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
      241 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
      301 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
      361 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
      421 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
      481 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
      541 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
      601 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
      661 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
      721 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
      781 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
      841 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
      901 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
      961 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     1021 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     1081 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     1141 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     1201 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     1261 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     1321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     1441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1801 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1861 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1921 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     1981 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     2041 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     2101 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     2161 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2221 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     2281 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2341 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     2401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2461 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2581 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     2641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     2701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2761 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2821 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     2881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3001 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     3061 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     3121 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     3181 tgatttaaat ggtcagttag agcggccgcc ccacacacca tagcttcaaa atgtttctac
     3241 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     3301 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     3361 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     3421 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct
     3481 ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt ctcaagtttc
     3541 agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc
     3601 atagcaatct aatctaagtt ttaattacaa aatggcagtg aaaagaacaa tcaagggtac
     3661 acaccactgg ttgttactca caattttgac ttcactttta gttttagtcc agagtactca
     3721 atggtctttg ttcttttttt tgtatgaaaa tgccttttta ccttttgcta tgggtattat
     3781 tgctatgtct gcttttgcaa tgatgtttgt caaacataag catgcatttc tctgtttgtt
     3841 tttgttacct tctcttgcca ctgtagctta ttttaatatg gtctatatgc ctgctagttg
     3901 ggtgatgcgt attatgacat ggttggatat ggttgatact agtttgtctg gttttaagct
     3961 aaaagactgt gttatgtatg catcagctgt agtgttacta atccttatga cagcaagaac
     4021 tgtgtatgat gatggtgcta ggagagtgtg gacacttatg aatgtcttga cactcgttta
     4081 taaagtttat tatggtaatg ctttagatca agccatttcc atgtgggctc ttataatctc
     4141 tgttacttct aactactcag gtgtagttac aactgtcatg tttttggcca gaggtattgt
     4201 ttttatgtgt gttgagtatt gccctatttt cttcataact ggtaatacac ttcagtgtat
     4261 aatgctagtt tattgtttct taggctattt ttgtacttgt tactttggcc tcttttgttt
     4321 actcaaccgc tactttagac tgactcttgg tgtttatgat tacttagttt ctacacagga
     4381 gtttagatat atgaattcac agggactact cccacccaag aatagcatag atgccttcaa
     4441 actcaacatt aaattgttgg gtgttggtgg caaaccttgt atcaaagtag ccactgtact
     4501 cgtgccgaga ggttctaaaa tcgaggaagg taagctagtc atttggataa atggagataa
     4561 gggttacaac ggtttggcag aagtaggcaa gaaatttgaa aaggacacag gtatcaaggt
     4621 gacggttgaa catccagaca agttagaaga aaaattccct caagttgctg ctactggtga
     4681 tggtcccgac attattttct gggcccacga tcgttttggt ggatatgctc agtcaggttt
     4741 gctggctgag atcaccccag acaaggcctt ccaagataaa ttgtacccat tcacttggga
     4801 tgctgtcaga tacaacggta agcttatagc atatccaatt gctgttgaag ccttatcgtt
     4861 gatctacaat aaggacttgt tacctaaccc accaaagaca tgggaagaaa ttccagcttt
     4921 ggataaagag ctaaaggcca agggaaaaag cgctttgatg tttaacttac aagaacctta
     4981 tttcacctgg ccattaatcg ctgctgacgg tggttacgca tttaaatacg aaaacggtaa
     5041 gtatgacatt aaggatgtcg gcgttgataa tgctggtgca aaggctggtt tgaccttctt
     5101 ggttgattta ataaagaata aacacatgaa cgctgacact gactactcta tcgccgaagc
     5161 cgctttcaac aagggtgaaa ctgctatgac cataaatggt ccatgggcct ggagtaacat
     5221 tgatacttct aaagtaaact atggtgtcac tgttttgcct acattcaaag gccaaccatc
     5281 caagcccttt gtcggtgttc tatccgctgg catcaatgct gcttctccaa acaaggagct
     5341 agctaaagaa tttttggaaa attacttgtt aactgacgaa ggtttggagg ccgttaacaa
     5401 ggacaagcca ttgggtgctg tcgctttgaa atcttacgaa gaagaattgg ctaaggatcc
     5461 cagaattgcc gctaccatgg agaacgccca aaagggtgaa attatgccaa acatcccaca
     5521 aatgtcagcc ttctggtacg cagttaggac tgccgtgatt aatgctgctt ccggtagaca
     5581 aaccgtcgac gaagcattaa aagatgctca aacgtaaaca ggcccctttt cctttgtcga
     5641 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac
     5701 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt
     5761 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg
     5821 gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta
     5881 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     5941 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     6001 aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
     6061 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
     6121 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
     6181 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
     6241 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac
     6301 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
     6361 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
     6421 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
     6481 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
     6541 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
     6601 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat
     6661 gcattgggtc aacagtatag aaccgtggat gatgtggttt cta
//
