LOCUS       sequence-297591-        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"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             683..700
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1843..1861)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2212..3015
                     /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      3098..4080
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4158..4187
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4209..4796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4808..4835)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4859..5050)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5051..5842,5843..5911))
                     /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     5674..5693
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5912..6014)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6015..6046)
                     /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 aactcgtgcc gagaggttct aaaatcgagg aaggtaagct
      721 agtcatttgg ataaatggag ataagggtta caacggtttg gcagaagtag gcaagaaatt
      781 tgaaaaggac acaggtatca aggtgacggt tgaacatcca gacaagttag aagaaaaatt
      841 ccctcaagtt gctgctactg gtgatggtcc cgacattatt ttctgggccc acgatcgttt
      901 tggtggatat gctcagtcag gtttgctggc tgagatcacc ccagacaagg ccttccaaga
      961 taaattgtac ccattcactt gggatgctgt cagatacaac ggtaagctta tagcatatcc
     1021 aattgctgtt gaagccttat cgttgatcta caataaggac ttgttaccta acccaccaaa
     1081 gacatgggaa gaaattccag ctttggataa agagctaaag gccaagggaa aaagcgcttt
     1141 gatgtttaac ttacaagaac cttatttcac ctggccatta atcgctgctg acggtggtta
     1201 cgcatttaaa tacgaaaacg gtaagtatga cattaaggat gtcggcgttg ataatgctgg
     1261 tgcaaaggct ggtttgacct tcttggttga tttaataaag aataaacaca tgaacgctga
     1321 cactgactac tctatcgccg aagccgcttt caacaagggt gaaactgcta tgaccataaa
     1381 tggtccatgg gcctggagta acattgatac ttctaaagta aactatggtg tcactgtttt
     1441 gcctacattc aaaggccaac catccaagcc ctttgtcggt gttctatccg ctggcatcaa
     1501 tgctgcttct ccaaacaagg agctagctaa agaatttttg gaaaattact tgttaactga
     1561 cgaaggtttg gaggccgtta acaaggacaa gccattgggt gctgtcgctt tgaaatctta
     1621 cgaagaagaa ttggctaagg atcccagaat tgccgctacc atggagaacg cccaaaaggg
     1681 tgaaattatg ccaaacatcc cacaaatgtc agccttctgg tacgcagtta ggactgccgt
     1741 gattaatgct gcttccggta gacaaaccgt cgacgaagca ttaaaagatg ctcaaacgta
     1801 aacaggcccc ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg
     1861 ccctcctccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     1921 ccctatttat tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt
     1981 tttctttttt ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt
     2041 ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac
     2101 agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta
     2161 ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa
     2221 gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat
     2281 atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa
     2341 ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat
     2401 atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag
     2461 tacaattttt tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg
     2521 cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt
     2581 gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa
     2641 cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa
     2701 tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt
     2761 gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt
     2821 gtgggtttag atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg
     2881 gtttctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat
     2941 gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc
     3001 ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt
     3061 agagcttcaa tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt
     3121 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     3181 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     3241 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     3301 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     3361 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
     3421 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
     3481 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
     3541 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
     3601 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
     3661 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
     3721 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
     3781 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
     3841 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
     3901 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
     3961 agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa
     4021 agtgcgtcat cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag
     4081 ctagagaata ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg
     4141 actgcaatca actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa
     4201 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4261 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa
     4321 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4681 cgagggagct tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4741 tctgacttga gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4801 ccagcaacgc agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc
     4861 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     4921 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     4981 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     5041 gtttggtcac ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5101 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac
     5161 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5221 cagcaataaa ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg
     5281 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5341 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5401 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5461 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag
     5521 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga
     5581 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5641 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5701 taaaagtgct catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc
     5761 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5821 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5881 taagggcgac acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg
     5941 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     6001 ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg
     6061 tcaggtatga tttaaatggt cagttaga
//
