LOCUS       sequence-297868-        6178 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 6178)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6178)
  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-m4046297
FEATURES             Location/Qualifiers
     source          1..6178
                     /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"
     primer_bind     complement(1933..1951)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2302..3105
                     /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      3188..4170
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4248..4277
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4299..4886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4898..4925)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4949..5140)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5141..5932,5933..6001))
                     /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     5764..5783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6002..6104)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6105..6136)
                     /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 tttnnnnntc 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 aaaatcgagg aaggtaagct agtcatttgg ataaatggag ataagggtta
      841 caacggtttg gcagaagtag gcaagaaatt tgaaaaggac acaggtatca aggtgacggt
      901 tgaacatcca gacaagttag aagaaaaatt ccctcaagtt gctgctactg gtgatggtcc
      961 cgacattatt ttctgggccc acgatcgttt tggtggatat gctcagtcag gtttgctggc
     1021 tgagatcacc ccagacaagg ccttccaaga taaattgtac ccattcactt gggatgctgt
     1081 cagatacaac ggtaagctta tagcatatcc aattgctgtt gaagccttat cgttgatcta
     1141 caataaggac ttgttaccta acccaccaaa gacatgggaa gaaattccag ctttggataa
     1201 agagctaaag gccaagggaa aaagcgcttt gatgtttaac ttacaagaac cttatttcac
     1261 ctggccatta atcgctgctg acggtggtta cgcatttaaa tacgaaaacg gtaagtatga
     1321 cattaaggat gtcggcgttg ataatgctgg tgcaaaggct ggtttgacct tcttggttga
     1381 tttaataaag aataaacaca tgaacgctga cactgactac tctatcgccg aagccgcttt
     1441 caacaagggt gaaactgcta tgaccataaa tggtccatgg gcctggagta acattgatac
     1501 ttctaaagta aactatggtg tcactgtttt gcctacattc aaaggccaac catccaagcc
     1561 ctttgtcggt gttctatccg ctggcatcaa tgctgcttct ccaaacaagg agctagctaa
     1621 agaatttttg gaaaattact tgttaactga cgaaggtttg gaggccgtta acaaggacaa
     1681 gccattgggt gctgtcgctt tgaaatctta cgaagaagaa ttggctaagg atcccagaat
     1741 tgccgctacc atggagaacg cccaaaaggg tgaaattatg ccaaacatcc cacaaatgtc
     1801 agccttctgg tacgcagtta ggactgccgt gattaatgct gcttccggta gacaaaccgt
     1861 cgacgaagca ttaaaagatg ctcaaacgta aacaggcccc ttttcctttg tcgatatcat
     1921 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
     1981 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
     2041 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc
     2101 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
     2161 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     2221 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
     2281 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     2341 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     2401 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     2461 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     2521 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     2581 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     2641 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     2701 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     2761 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     2821 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     2881 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     2941 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     3001 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     3061 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     3121 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     3181 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     3241 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     3301 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     3361 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     3421 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     3481 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     3541 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     3601 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     3661 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     3721 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     3781 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     3841 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     3901 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     3961 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     4021 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     4081 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca
     4141 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc
     4201 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac
     4261 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc
     4321 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4381 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata
     4441 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4501 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4561 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4621 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4681 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4741 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt
     4801 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct
     4861 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt
     4921 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg
     4981 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     5041 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     5101 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg
     5161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5221 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc
     5281 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg
     5341 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5401 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5461 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5521 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5581 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5641 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa
     5701 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5761 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5821 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5881 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5941 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6001 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     6061 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     6121 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
