LOCUS       sequence-298472-        5593 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 5593)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5593)
  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-m4046689
FEATURES             Location/Qualifiers
     source          1..5593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1348..1366)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1717..2520
                     /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      2603..3585
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3663..3692
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3714..4301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4313..4340)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4364..4555)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4556..5347,5348..5416))
                     /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     5179..5198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5417..5519)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5520..5551)
                     /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 caataaacag gccccttttc
     1321 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     1381 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1441 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     1501 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     1561 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     1621 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     1681 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     1741 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     1801 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     1861 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     1921 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     1981 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     2041 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     2101 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     2161 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     2221 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     2281 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     2341 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     2401 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     2461 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     2521 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     2581 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     2641 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2701 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     2761 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2821 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     2881 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     2941 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     3001 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     3061 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     3121 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     3181 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     3241 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     3301 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     3361 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     3421 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     3481 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     3541 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     3601 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     3661 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     3721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3781 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     3841 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3901 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3961 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4021 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4081 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4141 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4201 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     4261 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     4321 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     4381 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4441 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4501 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     4561 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4621 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     4681 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4741 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     4801 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4861 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4921 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4981 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     5041 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     5101 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5161 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5221 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     5281 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5341 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5401 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     5461 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     5521 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     5581 atggtcagtt aga
//
