LOCUS       sequence-287623-        5104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5104)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5104)
  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-m4046421
FEATURES             Location/Qualifiers
     source          1..5104
                     /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             773..790
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             791..814
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(859..877)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1228..2031
                     /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      2114..3096
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3174..3203
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3225..3812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3824..3851)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3875..4066)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4067..4858,4859..4927))
                     /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     4690..4709
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4928..5030)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5031..5062)
                     /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 atgaacaatg agctttctcc tgttgcttta cggcagatgt cctgcgcagc
      481 tggaacaacg caaactgcct gtactgacga taacgctcta gcttactata acaccactaa
      541 aggtggtaga ttcgtcttgg ccttgttgtc agacctgcaa gatttgaagt gggcgagatt
      601 tccaaagagc gatggcactg gtaccatata caccgaatta gaaccaccct gtcgtttcgt
      661 gaccgacact ccaaaaggtc cgaaggttaa gtatttgtac tttattaagg gtttaaataa
      721 cttgaatcga ggtatggttc tggggtcttt ggctgctaca gtcagactcc aactcgtgcc
      781 gagaggttct tggagccatc cgcagtttga aaaataaaca ggcccctttt cctttgtcga
      841 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac
      901 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt
      961 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg
     1021 gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta
     1081 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     1141 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     1201 aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
     1261 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
     1321 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
     1381 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
     1441 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac
     1501 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
     1561 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
     1621 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
     1681 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
     1741 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
     1801 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat
     1861 gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt
     1921 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     1981 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     2041 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     2101 ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     2161 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     2221 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2281 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2341 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2401 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2461 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     2521 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     2581 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     2641 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     2701 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     2761 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     2821 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     2881 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     2941 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     3001 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg
     3061 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc
     3121 ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc actggctcac
     3181 cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt
     3241 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3301 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac
     3361 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3421 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3481 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3541 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3601 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3661 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg
     3721 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt
     3781 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc
     3841 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga
     3901 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3961 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4021 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa
     4081 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4141 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga
     4201 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga
     4261 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt
     4321 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4381 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4441 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4501 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4561 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt
     4621 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4681 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4741 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4801 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4861 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4921 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     4981 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc
     5041 ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt
     5101 taga
//
