LOCUS       sequence-287607-        5341 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 5341)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5341)
  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-m4046461
FEATURES             Location/Qualifiers
     source          1..5341
                     /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             968..985
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             986..1051
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1096..1114)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1465..2268
                     /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      2351..3333
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3411..3440
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3462..4049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4061..4088)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4112..4303)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4304..5095,5096..5164))
                     /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     4927..4946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5165..5267)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5268..5299)
                     /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 atggagagcc ttgtccctgg tttcaacgag aaaacacacg tccaactcag
      481 tttgcctgtt ttacaggttc gcgacgtgct cgtacgtggc tttggagact ccgtggagga
      541 ggtcttatca gaggcacgtc aacatcttaa agatggcact tgtggcttag tagaagttga
      601 aaaaggcgtt ttgcctcaac ttgaacagcc ctatgtgttc atcaaacgtt cggatgctcg
      661 aactgcacct catggtcatg ttatggttga gctggtagca gaactcgaag gcattcagta
      721 cggtcgtagt ggtgagacac ttggtgtcct tgtccctcat gtgggcgaaa taccagtggc
      781 ttaccgcaag gttcttcttc gtaagaacgg taataaagga gctggtggcc atagttacgg
      841 cgccgatcta aagtcatttg acttaggcga cgagcttggc actgatcctt atgaagattt
      901 tcaagaaaac tggaacacta aacatagcag tggtgttacc cgtgaactca tgcgtgagct
      961 taacggactc gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca
     1021 tgatattgat tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat
     1081 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     1141 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     1201 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     1261 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     1321 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     1381 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     1441 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     1501 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     1561 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     1621 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     1681 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     1741 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     1801 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     1861 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     1921 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     1981 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     2041 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     2101 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     2161 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     2221 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     2281 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     2341 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     2401 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     2461 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     2521 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2581 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     2641 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     2701 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     2761 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     2821 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     2881 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     2941 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     3001 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     3061 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     3121 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     3181 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     3241 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     3301 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     3361 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     3421 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     3481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3541 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     3601 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3661 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3721 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3781 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3841 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3901 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     3961 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     4021 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     4081 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     4141 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     4201 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4261 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     4321 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4381 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     4441 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     4501 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4561 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4621 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4681 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4741 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4801 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     4861 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4921 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4981 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5041 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5101 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5161 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     5221 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     5281 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     5341 a
//
