LOCUS       sequence-300484-        4966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4966)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4966)
  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-m4046627
FEATURES             Location/Qualifiers
     source          1..4966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(258..1061)
                     /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"
     primer_bind     1412..1430
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(1734..2147)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      2193..2224
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2225..2327
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2328..2396,2397..3188)
                     /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     complement(2546..2565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3189..3380
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      3404..3431
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3443..4030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4052..4081)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(4159..4966,1..175)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
       61 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
      121 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttactcg
      181 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
      241 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
      301 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
      361 ccaacaataa taatgtcaga tcctgtagaa accacatcat ccacggttct atactgttga
      421 cccaatgcat ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
      481 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
      541 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
      601 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
      661 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
      721 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
      781 aattttctgt cctcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
      841 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
      901 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
      961 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     1021 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     1081 cacgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     1141 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     1201 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgtg tactgcagtg cactagtggt
     1261 tcggcggccg ctttgtacag aaaaaaaaga aaaatttgaa atataaataa cgttcttaat
     1321 actaacataa ctattaaaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt
     1381 ccttttcggt tagagcggat gtgggaggag ggcgtgaatg taagcgtgac ataactaatt
     1441 acatgatatc gacaaaggaa aaggggcctg tttataaggt tgccctgttg tccagcattt
     1501 cttcacaaag cttgtttatg tctacagcac cctgcatgga aagcaaaaca gaaagtagtg
     1561 aaaccatttt ttcaaaggct tcagtagtat ctttagctaa gagaatgtca ttgtgtaact
     1621 ggacacattg agcccacaat ttagatgatg attctactct gagttgttgc aaaactgaga
     1681 gtaagactac tgatgtgcac tttacatctg acattttcat tttgtaatta aaacttagat
     1741 tagattgcta tgctttcttt ctaatgagca agaagtaaaa aaagttgtaa tagaacaaga
     1801 aaaatgaaac tgaaacttga gaaattgaag accgtttatt aacttaaata tcaatgggag
     1861 gtcatcgaaa gagaaaaaaa tcaaaaaaaa aaattttcaa gaaaaagaaa cgtgataaaa
     1921 atttttattg cctttttcga cgaagaaaaa gaaacgaggc ggtctctttt ttcttttcca
     1981 aacctttagt acgggtaatt aacgacaccc tagaggaaga aagaggggaa atttagtatg
     2041 ctgtgcttgg gtgttttgaa gtggtacggc gatgcgcgga gtccgagaaa atctggaaga
     2101 gtaaaaaagg agtagaaaca ttttgaagct atggtgtgtg gggcggccgc tctaactgac
     2161 catttaaatc atacctgacc tccatagcag aaagtcaaaa gcctccgacc ggaggctttt
     2221 gacttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     2281 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg agtattcaac
     2341 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2401 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2461 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt acgccccgaa gaacgttttc
     2521 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2581 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2641 cagtcacaga aaagcatctc acggatggca tgacagtaag agaattatgc agtgctgcca
     2701 taaccatgag tgataacact gcggccaact tacttctggc aacgatcgga ggaccgaagg
     2761 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2821 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2881 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2941 taatagactg gatggaggcg gataaagttg caggatcact tctgcgctcg gccctcccgg
     3001 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3061 cagcactggg gccagatggt aagccctccc gcatcgtagt tatctacacg acggggagtc
     3121 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3181 attggtaagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
     3241 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
     3301 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
     3361 atgacggtga aaacctctga tgagggccca aatgtaatca cctggctcac cttcgggtgg
     3421 gcctttctgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3481 atcgatgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttta
     3541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3901 gcgctctgct gaagccagtt acctcggaaa aagagttggt agctcttgat ccggcaaaca
     3961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4021 aggatctcaa gaagatcctt tgattttcta ccgaagaaag gcccacccgt gaaggtgagc
     4081 cagtgagttg attgcagtcc agttacgctg gagtcaagct tgtaacacgc ggccgctcgg
     4141 gaagttccta ttctctagct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa
     4201 gcgctctgat gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa
     4261 aatattgcga ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc
     4321 tgtgctatat ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa
     4381 ctcgacctct acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta
     4441 gtaagaacta ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg
     4501 tagtatatag agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat
     4561 caacgctatc actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc
     4621 ggggatgcct ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg
     4681 gaagtggagt caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta
     4741 accttaacgg acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag
     4801 gagaaaaaaa gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt
     4861 gtagaacaaa aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt
     4921 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagc
//
