LOCUS       sequence-297676-        6082 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 6082)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6082)
  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-m4046423
FEATURES             Location/Qualifiers
     source          1..6082
                     /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             677..694
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1837..1855)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2206..3009
                     /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      3092..4074
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4152..4181
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4203..4790
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4802..4829)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4853..5044)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5045..5836,5837..5905))
                     /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     5668..5687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5906..6008)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6009..6040)
                     /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 atgaaaatgt cagatgtaaa gtgcacatca gtagtcttac tctcagtttt
      481 gcaacaactc agagtagaat catcatctaa attgtgggct caatgtgtcc agttacacaa
      541 tgacattctc ttagctaaag atactactga agcctttgaa aaaatggttt cactactttc
      601 tgttttgctt tccatgcagg gtgctgtaga cataaacaag ctttgtgaag aaatgctgga
      661 caacagggca accttactcg tgccgagagg ttctaaaatc gaggaaggta agctagtcat
      721 ttggataaat ggagataagg gttacaacgg tttggcagaa gtaggcaaga aatttgaaaa
      781 ggacacaggt atcaaggtga cggttgaaca tccagacaag ttagaagaaa aattccctca
      841 agttgctgct actggtgatg gtcccgacat tattttctgg gcccacgatc gttttggtgg
      901 atatgctcag tcaggtttgc tggctgagat caccccagac aaggccttcc aagataaatt
      961 gtacccattc acttgggatg ctgtcagata caacggtaag cttatagcat atccaattgc
     1021 tgttgaagcc ttatcgttga tctacaataa ggacttgtta cctaacccac caaagacatg
     1081 ggaagaaatt ccagctttgg ataaagagct aaaggccaag ggaaaaagcg ctttgatgtt
     1141 taacttacaa gaaccttatt tcacctggcc attaatcgct gctgacggtg gttacgcatt
     1201 taaatacgaa aacggtaagt atgacattaa ggatgtcggc gttgataatg ctggtgcaaa
     1261 ggctggtttg accttcttgg ttgatttaat aaagaataaa cacatgaacg ctgacactga
     1321 ctactctatc gccgaagccg ctttcaacaa gggtgaaact gctatgacca taaatggtcc
     1381 atgggcctgg agtaacattg atacttctaa agtaaactat ggtgtcactg ttttgcctac
     1441 attcaaaggc caaccatcca agccctttgt cggtgttcta tccgctggca tcaatgctgc
     1501 ttctccaaac aaggagctag ctaaagaatt tttggaaaat tacttgttaa ctgacgaagg
     1561 tttggaggcc gttaacaagg acaagccatt gggtgctgtc gctttgaaat cttacgaaga
     1621 agaattggct aaggatccca gaattgccgc taccatggag aacgcccaaa agggtgaaat
     1681 tatgccaaac atcccacaaa tgtcagcctt ctggtacgca gttaggactg ccgtgattaa
     1741 tgctgcttcc ggtagacaaa ccgtcgacga agcattaaaa gatgctcaaa cgtaaacagg
     1801 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     1861 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     1921 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     1981 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     2041 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     2101 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     2161 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     2221 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     2281 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     2341 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     2401 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     2461 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     2521 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     2581 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     2641 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     2701 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     2761 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     2821 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     2881 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     2941 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     3001 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     3061 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     3121 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     3181 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     3241 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3301 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     3361 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     3421 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     3481 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     3541 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     3601 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     3661 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     3721 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     3781 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     3841 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     3901 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     3961 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     4021 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     4081 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     4141 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     4201 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4261 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     4321 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4381 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4441 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4501 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4561 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4621 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4681 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4741 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4801 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     4861 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     4921 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     4981 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     5041 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5101 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     5161 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5221 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     5281 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5341 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5401 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5461 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     5521 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     5581 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5641 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5701 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     5761 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5821 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5881 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     5941 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     6001 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     6061 atgatttaaa tggtcagtta ga
//
