LOCUS       sequence-287654-        5344 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 5344)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5344)
  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-m4046586
FEATURES             Location/Qualifiers
     source          1..5344
                     /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..1054
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1099..1117)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1468..2271
                     /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      2354..3336
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3414..3443
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3465..4052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4064..4091)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4115..4306)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4307..5098,5099..5167))
                     /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     4930..4949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5168..5270)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5271..5302)
                     /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 gaggttctga ttataaagat catgatggtg attataaaga
     1021 tcatgatatt gattataaag atgatgatga taaataaaca ggcccctttt cctttgtcga
     1081 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac
     1141 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt
     1201 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg
     1261 gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta
     1321 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     1381 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     1441 aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
     1501 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
     1561 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
     1621 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
     1681 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac
     1741 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
     1801 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
     1861 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
     1921 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
     1981 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
     2041 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat
     2101 gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt
     2161 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     2221 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     2281 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     2341 ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     2401 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     2461 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2521 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2581 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2641 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2701 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     2761 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     2821 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     2881 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     2941 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     3001 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     3061 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     3121 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     3181 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     3241 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg
     3301 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc
     3361 ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc actggctcac
     3421 cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt
     3481 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3541 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac
     3601 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3841 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg
     3961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt
     4021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc
     4081 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga
     4141 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     4201 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4261 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa
     4321 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4381 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga
     4441 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga
     4501 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt
     4561 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4621 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4681 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4741 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4801 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt
     4861 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4921 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4981 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5041 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5101 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5161 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     5221 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc
     5281 ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt
     5341 taga
//
