LOCUS       sequence-298477-        5260 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 5260)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5260)
  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-m4046654
FEATURES             Location/Qualifiers
     source          1..5260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1015..1033)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1384..2187
                     /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      2270..3252
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3330..3359
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3381..3968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3980..4007)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4031..4222)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4223..5014,5015..5083))
                     /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     4846..4865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5084..5186)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5187..5218)
                     /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 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     1021 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     1081 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     1141 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     1201 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     1261 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     1321 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     1381 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     1441 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     1501 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     1561 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     1621 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     1681 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     1741 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     1801 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     1861 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     1921 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     1981 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     2041 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     2101 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     2161 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     2221 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
     2281 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     2341 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     2401 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     2461 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa
     2521 caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct
     2581 aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga
     2641 taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc
     2701 tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg
     2761 ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc
     2821 atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa
     2881 cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg
     2941 ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta
     3001 gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg
     3061 gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc
     3121 aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt
     3181 tggttttttg aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc
     3241 tatactttct agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc
     3301 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     3361 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3421 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3481 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3541 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3601 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3661 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3721 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3781 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3841 caggagagcg cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg
     3901 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3961 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     4021 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     4081 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     4141 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     4201 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     4261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     4321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4381 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     4441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
     4681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4741 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
     4921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5041 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
     5101 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     5161 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
     5221 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
