LOCUS       sequence-291434-        6750 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 6750)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6750)
  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-m4046341
FEATURES             Location/Qualifiers
     source          1..6750
                     /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"
     CDS             1345..1362
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2505..2523)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2874..3677
                     /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      3760..4742
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4820..4849
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4871..5458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5470..5497)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5521..5712)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5713..6504,6505..6573))
                     /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     6336..6355
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6574..6676)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6677..6708)
                     /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 gcagtgaaaa gaacaatcaa gggtacacac cactggttgt tactcacaat tttgacttca
      541 cttttagttt tagtccagag tactcaatgg tctttgttct tttttttgta tgaaaatgcc
      601 tttttacctt ttgctatggg tattattgct atgtctgctt ttgcaatgat gtttgtcaaa
      661 cataagcatg catttctctg tttgtttttg ttaccttctc ttgccactgt agcttatttt
      721 aatatggtct atatgcctgc tagttgggtg atgcgtatta tgacatggtt ggatatggtt
      781 gatactagtt tgtctggttt taagctaaaa gactgtgtta tgtatgcatc agctgtagtg
      841 ttactaatcc ttatgacagc aagaactgtg tatgatgatg gtgctaggag agtgtggaca
      901 cttatgaatg tcttgacact cgtttataaa gtttattatg gtaatgcttt agatcaagcc
      961 atttccatgt gggctcttat aatctctgtt acttctaact actcaggtgt agttacaact
     1021 gtcatgtttt tggccagagg tattgttttt atgtgtgttg agtattgccc tattttcttc
     1081 ataactggta atacacttca gtgtataatg ctagtttatt gtttcttagg ctatttttgt
     1141 acttgttact ttggcctctt ttgtttactc aaccgctact ttagactgac tcttggtgtt
     1201 tatgattact tagtttctac acaggagttt agatatatga attcacaggg actactccca
     1261 cccaagaata gcatagatgc cttcaaactc aacattaaat tgttgggtgt tggtggcaaa
     1321 ccttgtatca aagtagccac tgtactcgtg ccgagaggtt ctaaaatcga ggaaggtaag
     1381 ctagtcattt ggataaatgg agataagggt tacaacggtt tggcagaagt aggcaagaaa
     1441 tttgaaaagg acacaggtat caaggtgacg gttgaacatc cagacaagtt agaagaaaaa
     1501 ttccctcaag ttgctgctac tggtgatggt cccgacatta ttttctgggc ccacgatcgt
     1561 tttggtggat atgctcagtc aggtttgctg gctgagatca ccccagacaa ggccttccaa
     1621 gataaattgt acccattcac ttgggatgct gtcagataca acggtaagct tatagcatat
     1681 ccaattgctg ttgaagcctt atcgttgatc tacaataagg acttgttacc taacccacca
     1741 aagacatggg aagaaattcc agctttggat aaagagctaa aggccaaggg aaaaagcgct
     1801 ttgatgttta acttacaaga accttatttc acctggccat taatcgctgc tgacggtggt
     1861 tacgcattta aatacgaaaa cggtaagtat gacattaagg atgtcggcgt tgataatgct
     1921 ggtgcaaagg ctggtttgac cttcttggtt gatttaataa agaataaaca catgaacgct
     1981 gacactgact actctatcgc cgaagccgct ttcaacaagg gtgaaactgc tatgaccata
     2041 aatggtccat gggcctggag taacattgat acttctaaag taaactatgg tgtcactgtt
     2101 ttgcctacat tcaaaggcca accatccaag ccctttgtcg gtgttctatc cgctggcatc
     2161 aatgctgctt ctccaaacaa ggagctagct aaagaatttt tggaaaatta cttgttaact
     2221 gacgaaggtt tggaggccgt taacaaggac aagccattgg gtgctgtcgc tttgaaatct
     2281 tacgaagaag aattggctaa ggatcccaga attgccgcta ccatggagaa cgcccaaaag
     2341 ggtgaaatta tgccaaacat cccacaaatg tcagccttct ggtacgcagt taggactgcc
     2401 gtgattaatg ctgcttccgg tagacaaacc gtcgacgaag cattaaaaga tgctcaaacg
     2461 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     2521 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     2581 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     2641 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     2701 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     2761 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     2821 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     2881 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     2941 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     3001 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     3061 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     3121 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     3181 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     3241 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     3301 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     3361 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     3421 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     3481 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     3541 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     3601 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     3661 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     3721 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     3781 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     3841 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     3901 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     3961 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     4021 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     4081 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     4141 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     4201 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     4261 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     4321 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     4381 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     4441 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     4501 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     4561 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     4621 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     4681 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     4741 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     4801 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     4861 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4921 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     4981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5341 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5401 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5461 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     5521 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     5581 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     5641 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     5701 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5761 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     5821 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5881 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     5941 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6001 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6061 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6121 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     6181 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6241 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6301 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6361 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     6421 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6481 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6541 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     6601 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     6661 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     6721 ggtcaggtat gatttaaatg gtcagttaga
//
