LOCUS       sequence-299931-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5296)
  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-m4046308
FEATURES             Location/Qualifiers
     source          1..5296
                     /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             971..988
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             989..1006
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1051..1069)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1420..2223
                     /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      2306..3288
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3366..3395
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3417..4004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4016..4043)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4067..4258)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4259..5050,5051..5119))
                     /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     4882..4901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5120..5222)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5223..5254)
                     /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 atggaatctt tggtccctgg atttaacgag aaaacccacg tacagctatc
      481 ccttcccgtg ttgcaagttc gtgacgttct ggttcggggt ttcggtgatt cagtcgaaga
      541 agtcttgagt gaagccaggc aacatttaaa ggatggtact tgtggcttag ttgaagtcga
      601 aaagggtgtg ttgccacaac ttgagcaacc atacgttttc attaagagat ctgacgcaag
      661 aacagcgcca cacggtcatg ttatggttga attggtcgct gaattggagg gtatacaata
      721 tggcagatcg ggtgaaactt taggtgtatt ggtcccacac gtaggtgaaa tccctgttgc
      781 ttacagaaaa gttttattaa gaaagaatgg taacaagggt gctggtggac actcctacgg
      841 agccgatctg aagtcttttg acttgggtga tgaattgggt actgatccat atgaagactt
      901 ccaagaaaat tggaacacca aacatagctc tggtgttacg cgtgaattga tgagagaact
      961 taatggtggt ctcgtgccga gaggttctca ccatcatcac catcattaaa caggcccctt
     1021 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac
     1081 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     1141 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     1201 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt
     1261 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     1321 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     1381 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     1441 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     1501 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     1561 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     1621 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     1681 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     1741 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     1801 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     1861 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     1921 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     1981 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     2041 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     2101 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     2161 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     2221 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     2281 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa
     2341 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     2401 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     2461 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2521 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     2581 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     2641 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2701 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2761 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2821 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     2881 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     2941 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     3001 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     3061 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     3121 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     3181 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     3241 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     3301 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     3361 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     3421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3481 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     3541 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3901 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3961 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     4021 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     4081 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     4141 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     4201 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     4261 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4321 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     4381 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4441 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     4501 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4561 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4621 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4681 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     4741 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     4801 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4861 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4921 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     4981 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5041 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5101 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     5161 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     5221 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     5281 taaatggtca gttaga
//
