LOCUS       sequence-291454-        6373 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 6373)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6373)
  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-m4046392
FEATURES             Location/Qualifiers
     source          1..6373
                     /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"
     primer_bind     complement(2128..2146)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2497..3300
                     /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      3383..4365
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4443..4472
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4494..5081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5093..5120)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5144..5335)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5336..6127,6128..6196))
                     /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     5959..5978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6197..6299)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6300..6331)
                     /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 gttctaaaat cgaggaaggt aagctagtca tttggataaa
     1021 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg
     1081 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc
     1141 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca
     1201 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt
     1261 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc
     1321 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat
     1381 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca
     1441 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga
     1501 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt
     1561 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat
     1621 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg
     1681 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg
     1741 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa
     1801 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc
     1861 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc
     1921 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa
     1981 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc
     2041 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc
     2101 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2161 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2221 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2281 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     2341 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     2401 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     2461 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     2521 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     2581 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     2641 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     2701 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     2761 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     2821 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     2881 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     2941 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3001 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     3061 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     3121 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     3181 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     3241 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     3301 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     3361 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     3421 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3481 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     3541 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3601 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     3661 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     3721 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     3781 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     3841 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     3901 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     3961 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4021 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     4081 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     4141 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     4201 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     4261 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     4321 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     4381 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     4441 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     4501 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4561 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     4621 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4681 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4741 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4801 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4861 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4921 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4981 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     5041 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     5101 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     5161 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     5221 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     5281 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     5341 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5401 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     5461 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5521 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     5581 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5641 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5701 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5761 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     5821 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     5881 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5941 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6001 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     6061 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6121 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6181 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     6241 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     6301 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     6361 atggtcagtt aga
//
