LOCUS       sequence-297616-        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"
     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"
     primer_bind     complement(2458..2476)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2827..3630
                     /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      3713..4695
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4773..4802
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4824..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5423..5450)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5474..5665)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5666..6457,6458..6526))
                     /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     6289..6308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6527..6629)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6630..6661)
                     /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 gttctaaaat
     1321 cgaggaaggt aagctagtca tttggataaa tggagataag ggttacaacg gtttggcaga
     1381 agtaggcaag aaatttgaaa aggacacagg tatcaaggtg acggttgaac atccagacaa
     1441 gttagaagaa aaattccctc aagttgctgc tactggtgat ggtcccgaca ttattttctg
     1501 ggcccacgat cgttttggtg gatatgctca gtcaggtttg ctggctgaga tcaccccaga
     1561 caaggccttc caagataaat tgtacccatt cacttgggat gctgtcagat acaacggtaa
     1621 gcttatagca tatccaattg ctgttgaagc cttatcgttg atctacaata aggacttgtt
     1681 acctaaccca ccaaagacat gggaagaaat tccagctttg gataaagagc taaaggccaa
     1741 gggaaaaagc gctttgatgt ttaacttaca agaaccttat ttcacctggc cattaatcgc
     1801 tgctgacggt ggttacgcat ttaaatacga aaacggtaag tatgacatta aggatgtcgg
     1861 cgttgataat gctggtgcaa aggctggttt gaccttcttg gttgatttaa taaagaataa
     1921 acacatgaac gctgacactg actactctat cgccgaagcc gctttcaaca agggtgaaac
     1981 tgctatgacc ataaatggtc catgggcctg gagtaacatt gatacttcta aagtaaacta
     2041 tggtgtcact gttttgccta cattcaaagg ccaaccatcc aagccctttg tcggtgttct
     2101 atccgctggc atcaatgctg cttctccaaa caaggagcta gctaaagaat ttttggaaaa
     2161 ttacttgtta actgacgaag gtttggaggc cgttaacaag gacaagccat tgggtgctgt
     2221 cgctttgaaa tcttacgaag aagaattggc taaggatccc agaattgccg ctaccatgga
     2281 gaacgcccaa aagggtgaaa ttatgccaaa catcccacaa atgtcagcct tctggtacgc
     2341 agttaggact gccgtgatta atgctgcttc cggtagacaa accgtcgacg aagcattaaa
     2401 agatgctcaa acgtaaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
     2461 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     2521 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     2581 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     2641 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     2701 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     2761 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     2821 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     2881 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     2941 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     3001 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     3061 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     3121 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     3181 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     3241 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     3301 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     3361 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     3421 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     3481 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     3541 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     3601 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     3661 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     3721 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     3781 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     3841 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     3901 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     3961 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     4021 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     4081 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     4141 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     4201 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     4261 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     4321 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     4381 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     4441 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     4501 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     4561 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     4621 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     4681 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     4741 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     4801 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4861 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4921 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     5461 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     5521 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     5581 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     5641 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     5701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5761 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5821 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     5881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     6121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6181 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     6361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     6541 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     6601 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     6661 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
