LOCUS       sequence-297816-        5640 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 5640)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5640)
  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-m4046692
FEATURES             Location/Qualifiers
     source          1..5640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(1395..1413)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1764..2567
                     /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      2650..3632
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3710..3739
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3761..4348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4360..4387)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4411..4602)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4603..5394,5395..5463))
                     /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     5226..5245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5464..5566)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5567..5598)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 tctgctgtca aaagaactat taagggtacc catcactggt tgctgctaac aatcttaact
      541 tccttgttag tgcttgttca aagtacccag tggtcgttgt ttttcttctt gtacgagaac
      601 gcctttctcc cattcgcaat ggggataatt gctatgtcag cttttgccat gatgttcgtt
      661 aagcacaaac atgcgttttt gtgcttattc ttgttgcctt ctctcgctac ggtagcttat
      721 ttcaatatgg tctacatgcc agcttcctgg gttatgcgta tcatgacttg gttggacatg
      781 gttgatacca gcctttctgg tttcaagttg aaagattgtg tcatgtacgc atctgccgtt
      841 gtattactaa ttcttatgac tgctagaacc gtctatgacg atggcgcgag aagggtttgg
      901 acactaatga acgtgttgac tttagtttac aaggtctact atggaaatgc cttggaccaa
      961 gctatctcca tgtgggcttt aataatctca gtcacttcta actacagtgg tgttgttact
     1021 acagtgatgt ttttggctag aggtattgtc ttcatgtgtg ttgaatattg tcccattttc
     1081 tttatcaccg gtaacacttt acaatgcatt atgttggttt actgtttctt aggttacttt
     1141 tgtacctgtt atttcggcct attttgtttg ttgaatcgat actttagact cacattgggt
     1201 gtttacgatt atttggtcag cactcaagaa tttagatata tgaactccca aggtttgtta
     1261 ccaccaaaga actctattga tgccttcaag ttgaacatca agctgttagg tgtcggtgga
     1321 aaaccttgca tcaaggtagc caccgtccaa taaacaggcc ccttttcctt tgtcgatatc
     1381 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     1441 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     1501 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     1561 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     1621 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     1681 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     1741 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     1801 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     1861 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     1921 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     1981 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     2041 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     2101 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     2161 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     2221 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     2281 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     2341 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     2401 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     2461 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     2521 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     2581 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     2641 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     2701 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     2761 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     2821 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2881 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     2941 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     3001 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     3061 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     3121 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     3181 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     3241 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     3301 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     3361 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     3421 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     3481 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     3541 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     3601 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     3661 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     3721 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     3781 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3841 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     3901 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3961 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4021 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4081 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4141 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4201 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     4261 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     4321 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     4381 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     4441 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     4501 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     4561 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     4621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4681 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     4741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     4801 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5041 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5101 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     5161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5281 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5461 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     5521 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     5581 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
