LOCUS       sequence-300443-        6088 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 6088)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6088)
  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-m4046590
FEATURES             Location/Qualifiers
     source          1..6088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(320..1123)
                     /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     1474..1492
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2635..2652)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(2918..3331)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      3377..3408
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3409..3511
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3512..3580,3581..4372)
                     /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(3730..3749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4373..4564
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4588..4615
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4627..5214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5236..5265)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(5343..6088,1..237)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta
       61 gcgctctcgc gttgcatttt tgttttacaa aaatgaagca cagattcttc gttggtaaaa
      121 tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa
      181 aattagcgct ctcgcgttgc atttttgttc tacaaaatga agcacagatg cttcgttact
      241 cgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat
      301 ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc
      361 ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca aatagtcctc
      421 ttccaacaat aataatgtca gatcctgtag aaaccacatc atccacggtt ctatactgtt
      481 gacccaatgc atctcccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt
      541 aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt
      601 cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc
      661 atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct
      721 gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta atgtctgccc
      781 attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag
      841 caaattttct gtcctcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct
      901 taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt agtaaacaaa
      961 ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg
     1021 aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac
     1081 taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc
     1141 tgcacgtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa
     1201 cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     1261 gttcggagat taccgaatca aaaaaatttc aaggaaaccg tgtactgcag tgcactagtg
     1321 gttcggcggc cgctttgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta
     1381 atactaacat aactattaaa aaaaataaat agggacctag acttcaggtt gtctaactcc
     1441 ttccttttcg gttagagcgg atgtgggagg agggcgtgaa tgtaagcgtg acataactaa
     1501 ttacatgata tcgacaaagg aaaaggggcc tgtttacgtt tgagcatctt ttaatgcttc
     1561 gtcgacggtt tgtctaccgg aagcagcatt aatcacggca gtcctaactg cgtaccagaa
     1621 ggctgacatt tgtgggatgt ttggcataat ttcacccttt tgggcgttct ccatggtagc
     1681 ggcaattctg ggatccttag ccaattcttc ttcgtaagat ttcaaagcga cagcacccaa
     1741 tggcttgtcc ttgttaacgg cctccaaacc ttcgtcagtt aacaagtaat tttccaaaaa
     1801 ttctttagct agctccttgt ttggagaagc agcattgatg ccagcggata gaacaccgac
     1861 aaagggcttg gatggttggc ctttgaatgt aggcaaaaca gtgacaccat agtttacttt
     1921 agaagtatca atgttactcc aggcccatgg accatttatg gtcatagcag tttcaccctt
     1981 gttgaaagcg gcttcggcga tagagtagtc agtgtcagcg ttcatgtgtt tattctttat
     2041 taaatcaacc aagaaggtca aaccagcctt tgcaccagca ttatcaacgc cgacatcctt
     2101 aatgtcatac ttaccgtttt cgtatttaaa tgcgtaacca ccgtcagcag cgattaatgg
     2161 ccaggtgaaa taaggttctt gtaagttaaa catcaaagcg ctttttccct tggcctttag
     2221 ctctttatcc aaagctggaa tttcttccca tgtctttggt gggttaggta acaagtcctt
     2281 attgtagatc aacgataagg cttcaacagc aattggatat gctataagct taccgttgta
     2341 tctgacagca tcccaagtga atgggtacaa tttatcttgg aaggccttgt ctggggtgat
     2401 ctcagccagc aaacctgact gagcatatcc accaaaacga tcgtgggccc agaaaataat
     2461 gtcgggacca tcaccagtag cagcaacttg agggaatttt tcttctaact tgtctggatg
     2521 ttcaaccgtc accttgatac ctgtgtcctt ttcaaatttc ttgcctactt ctgccaaacc
     2581 gttgtaaccc ttatctccat ttatccaaat gactagctta ccttcctcga ttttagaacc
     2641 tctcggcacg agttgtaaag tagctctgtt atctagcatt tcttcacaca acttgtttat
     2701 gtcaacagca ccttgcatag acaacaagac agacaacaaa gaaaccatct tttcgaaagc
     2761 ttcggttgta tctttggcca ataaaatatc attgtgcaat tggacgcatt gtgcccaaag
     2821 cttggaagag ctctccacac gtaactgttg tagaacactc aggagaacta ccgacgtaca
     2881 cttgacgtct gacattttgg acattttgta attaaaactt agattagatt gctatgcttt
     2941 ctttctaatg agcaagaagt aaaaaaagtt gtaatagaac aagaaaaatg aaactgaaac
     3001 ttgagaaatt gaagaccgtt tattaactta aatatcaatg ggaggtcatc gaaagagaaa
     3061 aaaatcaaaa aaaaaaattt tcaagaaaaa gaaacgtgat aaaaattttt attgcctttt
     3121 tcgacgaaga aaaagaaacg aggcggtctc ttttttcttt tccaaacctt tagtacgggt
     3181 aattaacgac accctagagg aagaaagagg ggaaatttag tatgctgtgc ttgggtgttt
     3241 tgaagtggta cggcgatgcg cggagtccga gaaaatctgg aagagtaaaa aaggagtaga
     3301 aacattttga agctatggtg tgtggggcgg ccgctctaac tgaccattta aatcatacct
     3361 gacctccata gcagaaagtc aaaagcctcc gaccggaggc ttttgacttg atcggcacgt
     3421 aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt
     3481 atcgagattt tcaggagcta aggaagctaa aatgagtatt caacatttcc gtgtcgccct
     3541 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3601 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3661 cagcggtaag atccttgaga gtttacgccc cgaagaacgt tttccaatga tgagcacttt
     3721 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3781 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3841 tctcacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3901 cactgcggcc aacttacttc tggcaacgat cggaggaccg aaggagctaa ccgctttttt
     3961 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4021 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4081 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4141 ggcggataaa gttgcaggat cacttctgcg ctcggccctc ccggctggct ggtttattgc
     4201 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4261 tggtaagccc tcccgcatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4321 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aagtgaccaa
     4381 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     4441 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     4501 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     4561 ctgatgaggg cccaaatgta atcacctggc tcaccttcgg gtgggccttt ctgcgttgct
     4621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgat gctcaagtca
     4681 gaggtggcga aacccgacag gactataaag ataccaggcg tttacccctg gaagctccct
     4741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5041 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5101 agttacctcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5161 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5221 cctttgattt tctaccgaag aaaggcccac ccgtgaaggt gagccagtga gttgattgca
     5281 gtccagttac gctggagtca agcttgtaac acgcggccgc tcgggaagtt cctattctct
     5341 agctagaaag tataggaact tcagagcgct tttgaaaacc aaaagcgctc tgatgacgca
     5401 ctttcaaaaa accaaaaacg caccggactg taacgagcta ctaaaatatt gcgaataccg
     5461 cttccacaaa cattgctcaa aagtatctct ttgctatata tctctgtgct atatccctat
     5521 ataacctacc catccacctt tcgctccttg aacttgcatc taaactcgac ctctacattt
     5581 tttatgttta tctctagtat tactctttag acaaaaaaat tgtagtaaga actattcata
     5641 gagtgaatcg aaaacaatac gaaaatgtaa acatttccta tacgtagtat atagagacaa
     5701 aatagaagaa accgttcata attttctgac caatgaagaa tcatcaacgc tatcactttc
     5761 tgttcacaaa gtatgcgcaa tccacatcgg tatagaatat aatcggggat gcctttatct
     5821 tgaaaaaatg cacccgcagc ttcgctagta atcagtaaac gcgggaagtg gagtcaggct
     5881 ttttttatgg aagagaaaat agacaccaaa gtagccttct tctaacctta acggacctac
     5941 agtgcaaaaa gttatcaaga gactgcatta tagagcgcac aaaggagaaa aaaagtaatc
     6001 taagatgctt tgttagaaaa atagcgctct cgggatgcat ttttgtagaa caaaaaagaa
     6061 gtatagattc tttgttggta aaatagcg
//
