LOCUS       sequence-287612-        5635 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 5635)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5635)
  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-m4046312
FEATURES             Location/Qualifiers
     source          1..5635
                     /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             1304..1321
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1322..1345
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1390..1408)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1759..2562
                     /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      2645..3627
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3705..3734
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3756..4343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4355..4382)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4406..4597)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4598..5389,5390..5458))
                     /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     5221..5240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5459..5561)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5562..5593)
                     /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 caactcgtgc cgagaggttc
     1321 ttggagccat ccgcagtttg aaaaataaac aggccccttt tcctttgtcg atatcatgta
     1381 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     1441 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     1501 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     1561 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     1621 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     1681 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     1741 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     1801 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     1861 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     1921 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     1981 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     2041 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     2101 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     2161 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     2221 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     2281 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     2341 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     2401 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     2461 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     2521 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     2581 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     2641 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     2701 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     2761 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     2821 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     2881 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     2941 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     3001 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     3061 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     3121 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     3181 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     3241 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     3301 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     3361 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     3421 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     3481 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     3541 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     3601 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
     3661 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     3721 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3841 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     3901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4201 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc
     4261 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     4321 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     4381 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     4441 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     4501 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4561 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     4621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4681 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4741 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     4801 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5041 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5101 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     5161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5281 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     5461 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     5521 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     5581 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
