LOCUS       sequence-291644-        4972 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 4972)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4972)
  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-m4046648
FEATURES             Location/Qualifiers
     source          1..4972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(727..745)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1096..1899
                     /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      1982..2964
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3042..3071
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3093..3680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3692..3719)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3743..3934)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3935..4726,4727..4795))
                     /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     4558..4577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4796..4898)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4899..4930)
                     /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 atgtccaaaa tgtcagacgt caagtgtacg tcggtagttc tcctgagtgt
      481 tctacaacag ttacgtgtgg agagctcttc caagctttgg gcacaatgcg tccaattgca
      541 caatgatatt ttattggcca aagatacaac cgaagctttc gaaaagatgg tttctttgtt
      601 gtctgtcttg ttgtctatgc aaggtgctgt tgacataaac aagttgtgtg aagaaatgct
      661 agataacaga gctactttac aataaacagg ccccttttcc tttgtcgata tcatgtaatt
      721 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
      781 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
      841 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca
      901 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
      961 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     1021 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     1081 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     1141 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     1201 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     1261 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     1321 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     1381 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     1441 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     1501 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     1561 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     1621 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     1681 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     1741 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     1801 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     1861 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     1921 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     1981 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     2041 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     2101 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     2161 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     2221 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     2281 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     2341 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     2401 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     2461 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     2521 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     2581 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     2641 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     2701 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     2761 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     2821 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     2881 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
     2941 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
     3001 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     3061 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3181 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
     3241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt
     3601 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     3661 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     3721 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3781 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3841 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3901 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac
     3961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4021 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca
     4141 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4381 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4441 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt
     4501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4621 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag
     4801 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     4861 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag
     4921 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
