LOCUS       sequence-300768-        5677 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 5677)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5677)
  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-m4046577
FEATURES             Location/Qualifiers
     source          1..5677
                     /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             1304..1321
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1322..1387
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1432..1450)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1801..2604
                     /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      2687..3669
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3747..3776
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3798..4385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4397..4424)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4448..4639)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4640..5431,5432..5500))
                     /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     5263..5282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5501..5603)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5604..5635)
                     /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 atgtctgctg tcaaaagaac tattaagggt acccatcact ggttgctgct
      481 aacaatctta acttccttgt tagtgcttgt tcaaagtacc cagtggtcgt tgtttttctt
      541 cttgtacgag aacgcctttc tcccattcgc aatggggata attgctatgt cagcttttgc
      601 catgatgttc gttaagcaca aacatgcgtt tttgtgctta ttcttgttgc cttctctcgc
      661 tacggtagct tatttcaata tggtctacat gccagcttcc tgggttatgc gtatcatgac
      721 ttggttggac atggttgata ccagcctttc tggtttcaag ttgaaagatt gtgtcatgta
      781 cgcatctgcc gttgtattac taattcttat gactgctaga accgtctatg acgatggcgc
      841 gagaagggtt tggacactaa tgaacgtgtt gactttagtt tacaaggtct actatggaaa
      901 tgccttggac caagctatct ccatgtgggc tttaataatc tcagtcactt ctaactacag
      961 tggtgttgtt actacagtga tgtttttggc tagaggtatt gtcttcatgt gtgttgaata
     1021 ttgtcccatt ttctttatca ccggtaacac tttacaatgc attatgttgg tttactgttt
     1081 cttaggttac ttttgtacct gttatttcgg cctattttgt ttgttgaatc gatactttag
     1141 actcacattg ggtgtttacg attatttggt cagcactcaa gaatttagat atatgaactc
     1201 ccaaggtttg ttaccaccaa agaactctat tgatgccttc aagttgaaca tcaagctgtt
     1261 aggtgtcggt ggaaaacctt gcatcaaggt agccaccgtc caactcgtgc cgagaggttc
     1321 tgattataaa gatcatgatg gtgattataa agatcatgat attgattata aagatgatga
     1381 tgataaataa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     1441 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     1501 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     1561 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     1621 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     1681 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     1741 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     1801 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     1861 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     1921 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     1981 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     2041 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     2101 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     2161 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     2221 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     2281 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     2341 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     2401 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     2461 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     2521 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     2581 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     2641 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     2701 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     2761 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     2821 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     2881 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     2941 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     3001 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     3061 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     3121 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     3181 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     3241 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     3301 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3361 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     3421 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     3481 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     3541 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     3601 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     3661 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     3721 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     3781 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3841 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3901 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3961 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4021 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4081 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4141 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4201 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4261 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     4321 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     4381 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     4441 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4501 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4561 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4621 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     4681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     4741 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4801 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     4861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4921 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     5101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5161 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5221 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     5341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5461 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     5521 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     5581 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     5641 ctatggaggt caggtatgat ttaaatggtc agttaga
//
