LOCUS       sequence-291523-        6376 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 6376)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6376)
  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-m4046568
FEATURES             Location/Qualifiers
     source          1..6376
                     /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"
     primer_bind     complement(2131..2149)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2500..3303
                     /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      3386..4368
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4446..4475
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4497..5084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5096..5123)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5147..5338)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5339..6130,6131..6199))
                     /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     5962..5981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6200..6302)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6303..6334)
                     /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 gaggttctaa aatcgaggaa ggtaagctag tcatttggat
     1021 aaatggagat aagggttaca acggtttggc agaagtaggc aagaaatttg aaaaggacac
     1081 aggtatcaag gtgacggttg aacatccaga caagttagaa gaaaaattcc ctcaagttgc
     1141 tgctactggt gatggtcccg acattatttt ctgggcccac gatcgttttg gtggatatgc
     1201 tcagtcaggt ttgctggctg agatcacccc agacaaggcc ttccaagata aattgtaccc
     1261 attcacttgg gatgctgtca gatacaacgg taagcttata gcatatccaa ttgctgttga
     1321 agccttatcg ttgatctaca ataaggactt gttacctaac ccaccaaaga catgggaaga
     1381 aattccagct ttggataaag agctaaaggc caagggaaaa agcgctttga tgtttaactt
     1441 acaagaacct tatttcacct ggccattaat cgctgctgac ggtggttacg catttaaata
     1501 cgaaaacggt aagtatgaca ttaaggatgt cggcgttgat aatgctggtg caaaggctgg
     1561 tttgaccttc ttggttgatt taataaagaa taaacacatg aacgctgaca ctgactactc
     1621 tatcgccgaa gccgctttca acaagggtga aactgctatg accataaatg gtccatgggc
     1681 ctggagtaac attgatactt ctaaagtaaa ctatggtgtc actgttttgc ctacattcaa
     1741 aggccaacca tccaagccct ttgtcggtgt tctatccgct ggcatcaatg ctgcttctcc
     1801 aaacaaggag ctagctaaag aatttttgga aaattacttg ttaactgacg aaggtttgga
     1861 ggccgttaac aaggacaagc cattgggtgc tgtcgctttg aaatcttacg aagaagaatt
     1921 ggctaaggat cccagaattg ccgctaccat ggagaacgcc caaaagggtg aaattatgcc
     1981 aaacatccca caaatgtcag ccttctggta cgcagttagg actgccgtga ttaatgctgc
     2041 ttccggtaga caaaccgtcg acgaagcatt aaaagatgct caaacgtaaa caggcccctt
     2101 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac
     2161 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     2221 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     2281 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt
     2341 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     2401 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     2461 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     2521 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     2581 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     2641 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     2701 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     2761 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     2821 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     2881 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     2941 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     3001 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     3061 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     3121 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     3181 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     3241 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     3301 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     3361 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa
     3421 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     3481 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     3541 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     3601 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     3661 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     3721 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     3781 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     3841 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     3901 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     3961 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     4021 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     4081 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     4141 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     4201 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     4261 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     4321 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     4381 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     4441 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     4501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4561 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     4621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4981 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5041 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     5101 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     5161 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     5221 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     5281 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     5341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5401 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     5461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5521 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     5581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5641 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5761 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     5821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     5881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5941 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6001 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     6061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6181 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     6241 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     6301 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     6361 taaatggtca gttaga
//
