LOCUS       sequence-287582-        5008 bp    DNA     circular SYN 15-APR-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 5008)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5008)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046244
FEATURES             Location/Qualifiers
     source          1..5008
                     /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"
     CDS             701..718
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(763..781)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1132..1935
                     /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      2018..3000
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3078..3107
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3129..3716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3728..3755)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3779..3970)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3971..4762,4763..4831))
                     /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     4594..4613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4832..4934)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4935..4966)
                     /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 caccatcatc accatcatta
      721 aacaggcccc ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg
      781 ccctcctccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
      841 ccctatttat tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt
      901 tttctttttt ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt
      961 ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac
     1021 agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta
     1081 ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa
     1141 gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat
     1201 atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa
     1261 ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat
     1321 atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag
     1381 tacaattttt tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg
     1441 cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt
     1501 gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa
     1561 cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa
     1621 tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt
     1681 gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt
     1741 gtgggtttag atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg
     1801 gtttctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat
     1861 gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc
     1921 ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt
     1981 agagcttcaa tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt
     2041 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     2101 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     2161 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     2221 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     2281 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
     2341 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
     2401 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
     2461 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
     2521 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
     2581 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
     2641 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
     2701 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
     2761 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
     2821 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
     2881 agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa
     2941 agtgcgtcat cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag
     3001 ctagagaata ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg
     3061 actgcaatca actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa
     3121 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3181 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa
     3241 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3301 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3361 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3421 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3481 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3541 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3601 cgagggagct tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3661 tctgacttga gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3721 ccagcaacgc agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc
     3781 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     3841 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3901 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3961 gtttggtcac ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac
     4081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4141 cagcaataaa ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg
     4201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag
     4441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga
     4501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4621 taaaagtgct catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc
     4681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4801 taagggcgac acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg
     4861 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     4921 ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg
     4981 tcaggtatga tttaaatggt cagttaga
//
