LOCUS       sequence-298101-        5140 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 5140)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5140)
  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-m4046379
FEATURES             Location/Qualifiers
     source          1..5140
                     /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             767..784
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             785..850
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(895..913)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1264..2067
                     /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
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFXXRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      2150..3132
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3210..3239
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3261..3848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3860..3887)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3911..4102)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4103..4894,4895..4963))
                     /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     4726..4745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4964..5066)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5067..5098)
                     /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 atgaatgagc ttagtcctgt tgcactacga cagatgtctt gtgctgccgg
      481 tactacacaa actgcttgca ctgatgacaa tgcgttagct tactacaaca caacaaaggg
      541 aggtaggttt gtacttgcac tgttatccga tttacaggat ttgaaatggg ctagattccc
      601 taagagtgat ggaactggta ctatctatac agaactggaa ccaccttgta ggtttgttac
      661 agacacacct aaaggtccta aagtgaagta tttatacttt attaaaggat taaacaacct
      721 aaatagaggt atggtacttg gtagtttagc tgccacagta cgtctactcg tgccgagagg
      781 ttctgattat aaagatcatg atggtgatta taaagatcat gatattgatt ataaagatga
      841 tgatgataaa taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
      901 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
      961 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     1021 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     1081 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     1141 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     1201 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     1261 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     1321 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     1381 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     1441 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     1501 ttatccgcca agtacaattt tttactcttc gnnnncagaa aatttgctga cattggtaat
     1561 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     1621 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     1681 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     1741 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     1801 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     1861 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     1921 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     1981 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     2041 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     2101 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
     2161 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     2221 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     2281 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     2341 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa
     2401 caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct
     2461 aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga
     2521 taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc
     2581 tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg
     2641 ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc
     2701 atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa
     2761 cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg
     2821 ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta
     2881 gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg
     2941 gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc
     3001 aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt
     3061 tggttttttg aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc
     3121 tatactttct agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc
     3181 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     3241 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3301 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3361 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3421 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3481 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3541 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3601 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3661 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3721 caggagagcg cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg
     3781 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3841 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     3901 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3961 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     4021 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     4081 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     4141 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     4201 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4261 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     4321 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4381 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4441 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4501 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
     4561 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4621 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4681 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4741 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
     4801 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4861 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4921 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
     4981 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     5041 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
     5101 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
