LOCUS       sequence-298132-        5623 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 5623)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5623)
  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-m4046474
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gap             385..395
                     /estimated_length=11
     CDS             1298..1315
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1316..1333
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1378..1396)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1747..2550
                     /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      2633..3615
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3693..3722
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3744..4331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4343..4370)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4394..4585)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4586..5377,5378..5446))
                     /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     5209..5228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5447..5549)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5550..5581)
                     /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 tgctnnnnnn nnnnnaagca 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 gttctcacca
     1321 tcatcaccat cattaaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
     1381 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     1441 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     1501 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     1561 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     1621 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     1681 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     1741 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     1801 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     1861 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     1921 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     1981 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     2041 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     2101 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     2161 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     2221 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     2281 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     2341 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     2401 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     2461 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     2521 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     2581 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     2641 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     2701 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     2761 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     2821 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     2881 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     2941 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     3001 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     3061 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     3121 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     3181 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     3241 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     3301 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     3361 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     3421 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     3481 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     3541 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     3601 tcctatactn nnnagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     3661 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     3721 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3841 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4201 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     4261 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     4321 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     4381 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     4441 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     4501 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4561 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     4621 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4681 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4741 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     4801 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4861 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4921 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4981 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     5041 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5101 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5161 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5221 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     5281 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5341 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5401 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     5461 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     5521 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     5581 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
