LOCUS       sequence-300445-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7513)
  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-m4046641
FEATURES             Location/Qualifiers
     source          1..7513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..997
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      1075..1104
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1126..1713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1725..1752)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1776..1967)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1968..2759,2760..2828))
                     /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     2591..2610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2829..2931)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2932..2963)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3009..3422
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(6273..6291)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6642..7445
                     /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"
ORIGIN
        1 gttattaccc gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
       61 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
      121 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
      181 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
      241 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa
      301 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg
      361 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa
      421 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg
      481 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta
      541 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat
      601 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
      661 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
      721 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
      781 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
      841 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
      901 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg
      961 gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag
     1021 cggccgcgtg ttacaagctt gactccagcg taactggact gcaatcaact cactggctca
     1081 ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt
     1141 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1201 gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata
     1261 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1321 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1381 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1441 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1501 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1561 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac
     1621 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg
     1681 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc
     1741 cgaaggtgag ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg
     1801 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     1861 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     1921 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta
     1981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2041 cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg
     2101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccggg
     2161 agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2401 ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2461 gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag
     2521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2641 cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2821 atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     2881 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag
     2941 cctccggtcg gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag
     3001 ttagagcggc cgccccacac accatagctt caaaatgttt ctactccttt tttactcttc
     3061 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     3121 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     3181 aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     3241 tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
     3301 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
     3361 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
     3421 agttttaatt acaaaatggc tgatgcacaa tcgtttttaa accgggtttg cggtgtaagt
     3481 gcagcccgtc ttacaccgtg cggcacaggc actagtactg atgtcgtata cagggctttt
     3541 gacatctaca atgataaagt agctggtttt gctaaattcc taaaaactaa ttgttgtcgc
     3601 ttccaagaaa aggacgaaga tgacaattta attgattctt actttgtagt taagagacac
     3661 actttctcta actaccaaca tgaagaaaca atttataatt tacttaagga ttgtccagct
     3721 gttgctaaac atgacttctt taagtttaga atagacggtg acatggtacc acatatatca
     3781 cgtcaacgtc ttactaaata cacaatggca gacctcgtct atgctttaag gcattttgat
     3841 gaaggtaatt gtgacacatt aaaagaaata cttgtcacat acaattgttg tgatgatgat
     3901 tatttcaata aaaaggactg gtatgatttt gtagaaaacc cagatatatt acgcgtatac
     3961 gccaacttag gtgaacgtgt acgccaagct ttgttaaaaa cagtacaatt ctgtgatgcc
     4021 atgcgaaatg ctggtattgt tggtgtactg acattagata atcaagatct caatggtaac
     4081 tggtatgatt tcggtgattt catacaaacc acgccaggta gtggagttcc tgttgtagat
     4141 tcttattatt cattgttaat gcctatatta accttgacca gggctttaac tgcagagtca
     4201 catgttgaca ctgacttaac aaagccttac attaagtggg atttgttaaa atatgacttc
     4261 acggaagaga ggttaaaact ctttgaccgt tattttaaat attgggatca gacataccac
     4321 ccaaattgtg ttaactgttt ggatgacaga tgcattctgc attgtgcaaa ctttaatgtt
     4381 ttattctcta cagtgttccc acctacaagt tttggaccac tagtgagaaa aatatttgtt
     4441 gatggtgttc catttgtagt ttcaactgga taccacttca gagagctagg tgttgtacat
     4501 aatcaggatg taaacttaca tagctctaga cttagtttta aggaattact tgtgtatgct
     4561 gctgaccctg ctatgcacgc tgcttctggt aatctattac tagataaacg cactacgtgc
     4621 ttttcagtag ctgcacttac taacaatgtt gcttttcaaa ctgtcaaacc cggtaatttt
     4681 aacaaagact tctatgactt tgctgtgtct aagggtttct ttaaggaagg aagttctgtt
     4741 gaattaaaac acttcttctt tgctcaggat ggtaatgctg ctatcagcga ttatgactac
     4801 tatcgttata atctaccaac aatgtgtgat atcagacaac tactatttgt agttgaagtt
     4861 gttgataagt actttgattg ttacgatggt ggctgtatta atgctaacca agtcatcgtc
     4921 aacaacctag acaaatcagc tggttttcca tttaataaat ggggtaaggc tagactttat
     4981 tatgattcaa tgagttatga ggatcaagat gcacttttcg catatacaaa acgtaatgtc
     5041 atccctacta taactcaaat gaatcttaag tatgccatta gtgcaaagaa tagagctcgc
     5101 accgtagctg gtgtctctat ctgtagtact atgaccaata gacagtttca tcaaaaatta
     5161 ttgaaatcaa tagccgccac tagaggagct actgtagtaa ttggaacaag caaattctat
     5221 ggtggttggc acaacatgtt aaaaactgtt tatagtgatg tagaaaaccc tcaccttatg
     5281 ggttgggatt atcctaaatg tgatagagcc atgcctaaca tgcttagaat tatggcctca
     5341 cttgttcttg ctcgcaaaca tacaacgtgt tgtagcttgt cacaccgttt ctatagatta
     5401 gctaatgagt gtgctcaagt attgagtgaa atggtcatgt gtggcggttc actatatgtt
     5461 aaaccaggtg gaacctcatc aggagatgcc acaactgctt atgctaatag tgtttttaac
     5521 atttgtcaag ctgtcacggc caatgttaat gcacttttat ctactgatgg taacaaaatt
     5581 gccgataagt atgtccgcaa tttacaacac agactttatg agtgtctcta tagaaataga
     5641 gatgttgaca cagactttgt gaatgagttt tacgcatatt tgcgtaaaca tttctcaatg
     5701 atgatactct ctgacgatgc tgttgtgtgt ttcaatagca cttatgcatc tcaaggtcta
     5761 gtggctagca taaagaactt taagtcagtt ctttattatc aaaacaatgt ttttatgtct
     5821 gaagcaaaat gttggactga gactgacctt actaaaggac ctcatgaatt ttgctctcaa
     5881 catacaatgc tagttaaaca gggtgatgat tatgtgtacc ttccttaccc agatccatca
     5941 agaatcctag gggccggctg ttttgtagat gatatcgtaa aaacagatgg tacacttatg
     6001 attgaacggt tcgtgtcttt agctatagat gcttacccac ttactaaaca tcctaatcag
     6061 gagtatgctg atgtctttca tttgtactta caatacataa gaaagctaca tgatgagtta
     6121 acaggacaca tgttagacat gtattctgtt atgcttacta atgataacac ttcaaggtat
     6181 tgggaacctg agttttatga ggctatgtac acaccgcata cagtcttata aacaggcccc
     6241 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     6301 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     6361 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     6421 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     6481 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     6541 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     6601 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     6661 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     6721 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     6781 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     6841 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     6901 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     6961 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     7021 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     7081 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     7141 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     7201 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     7261 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     7321 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     7381 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     7441 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     7501 tttaattata tca
//
