LOCUS       sequence-287571-        5056 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 5056)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5056)
  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-m4046266
FEATURES             Location/Qualifiers
     source          1..5056
                     /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..766
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(811..829)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1180..1983
                     /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      2066..3048
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3126..3155
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3177..3764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3776..3803)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3827..4018)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4019..4810,4811..4879))
                     /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     4642..4661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4880..4982)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4983..5014)
                     /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 gattataaag atcatgatgg
      721 tgattataaa gatcatgata ttgattataa agatgatgat gataaataaa caggcccctt
      781 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac
      841 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
      901 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
      961 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt
     1021 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     1081 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     1141 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     1201 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     1261 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     1321 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     1381 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     1441 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     1501 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     1561 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     1621 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     1681 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     1741 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     1801 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     1861 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     1921 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     1981 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     2041 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa
     2101 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     2161 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     2221 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2281 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     2341 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     2401 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2461 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2521 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2581 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     2641 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     2701 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     2761 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     2821 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     2881 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     2941 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     3001 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     3061 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     3121 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     3181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3241 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     3301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3661 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3721 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     3781 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     3841 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3901 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3961 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     4021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4081 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     4141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4201 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     4261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4441 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     4501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     4561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4681 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     4741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4861 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     4921 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     4981 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     5041 taaatggtca gttaga
//
