LOCUS       sequence-300406-        7566 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 7566)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7566)
  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-m4046670
FEATURES             Location/Qualifiers
     source          1..7566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      999..1028
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1050..1637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1649..1676)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1700..1891)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1892..2683,2684..2752))
                     /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     2515..2534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2753..2855)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2856..2887)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2954..3395
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2954..3071
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3366..3389
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(6250..6268)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6619..7422
                     /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      join(7505..7566,1..921)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
       61 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
      121 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
      181 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
      241 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
      301 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
      361 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
      421 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
      481 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
      541 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
      601 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
      661 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
      721 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
      781 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
      841 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
      901 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
      961 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     1021 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1141 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1201 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1261 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1321 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1381 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1441 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1501 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     1561 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     1621 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     1681 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     1741 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     1801 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     1861 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     1921 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1981 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2041 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     2101 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2161 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2221 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2281 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2341 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2401 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     2461 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2521 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     2581 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2641 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2701 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     2761 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     2821 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct
     2881 tttgactttc tgctatggag gtcaggtatg atttaaatgg tcagttagag cggccgcttg
     2941 ctatattgaa gtacggatta gaagccgccg agcgggcgac agccctccga cggaagactc
     3001 tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
     3061 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
     3121 attggcagta acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg
     3181 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
     3241 tttttgatct attaacagat atataaatgg aaaagctgca taaccacttt aactaatact
     3301 ttcaacattt tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa
     3361 ttgttaatat acctctatac tttaacgtca aggagaaaaa actataatgt ccgccgatgc
     3421 acaatctttc ttaaatagag tctgcggcgt gagtgctgct cgtctaaccc cttgtggtac
     3481 gggtacttca acagacgtag tttacagggc gtttgatatt tataacgaca aggtcgctgg
     3541 tttcgctaaa tttttgaaga ccaactgttg tagattccag gagaaggatg aagacgacaa
     3601 tcttatcgat tcgtacttcg ttgttaaaag acatactttt tccaactatc aacacgaaga
     3661 aacaatatac aatttgctca aggactgtcc agccgtcgct aaacacgatt ttttcaagtt
     3721 cagaattgat ggggacatgg tgccacatat cagcagacaa cggttgacca agtacactat
     3781 ggctgattta gtttacgcct tgcgacactt tgacgaagga aactgtgata ctctgaagga
     3841 aattttggtt acctataact gctgtgacga tgattacttc aacaaaaagg actggtacga
     3901 cttcgtagag aatcccgaca tcttgagggt ttacgctaac ttaggtgaaa gagtcagaca
     3961 agctttgtta aagacagtcc aattttgcga tgctatgaga aatgcgggta ttgttggtgt
     4021 tttgactttg gataatcaag atttgaacgg taactggtat gacttcggtg attttatcca
     4081 gactaccccg ggttctgggg taccagttgt cgattcttac tattctctgt taatgccaat
     4141 actcacccta actagagcat tgactgccga atcccatgtt gacactgatt taactaagcc
     4201 atacattaaa tgggaccttc taaagtacga tttcaccgaa gaacgtttaa agttgttcga
     4261 cagatatttt aaatactggg accaaaccta ccaccctaat tgtgtcaact gtttggacga
     4321 cagatgtatt cttcactgcg ctaactttaa tgtgttgttc tcgactgttt ttccaccaac
     4381 ctctttcggt ccgctggttc gaaagatttt tgttgacggt gttccattcg tggtcagtac
     4441 gggatatcat ttcagagaat taggggtcgt tcataaccaa gacgtgaact tgcactcatc
     4501 aagattgtcc ttcaaggaat tgttggtcta cgcagctgat cctgcaatgc acgccgcttc
     4561 tggaaactta ttgctggata agagaacaac ctgtttcagc gttgccgctc ttactaataa
     4621 cgtcgcgttc caaactgtaa agccgggtaa ctttaataaa gacttctacg attttgctgt
     4681 tagcaagggt tttttcaaag aaggttctag tgttgagttg aaacactttt tctttgctca
     4741 agacggcaat gctgctatca gtgattatga ctattaccgt tacaacttac caacaatgtg
     4801 tgatatcaga cagttgcttt tcgtcgtcga agttgttgat aaatattttg actgttatga
     4861 tggtggttgc atcaatgcaa atcaagtcat tgtcaacaac ttagataagt ctgctggctt
     4921 cccattcaac aagtggggta aagccagatt gtactatgat tccatgtcct acgaggacca
     4981 agatgctttg ttcgcgtaca ccaagagaaa cgttattcct acaattactc aaatgaatct
     5041 aaagtacgct atctctgcaa aaaatcgggc cagaaccgtt gctggtgttt ctatctgttc
     5101 tacgatgact aaccgtcaat ttcatcagaa gctattgaaa tccatagctg ctactagagg
     5161 tgctactgta gtcatcggta cctcaaagtt ctatggtggt tggcacaata tgctcaagac
     5221 cgtttactct gatgtcgaaa acccacatct tatgggttgg gactacccta agtgtgatag
     5281 agccatgcca aatatgctaa gaattatggc atccttggtc ttggctagaa agcataccac
     5341 atgctgctca ttgtcccacc gtttttacag attggccaac gaatgtgctc aagtcctttc
     5401 tgaaatggtt atgtgtggtg gtagtctata tgtaaagcca ggcggaactt cgtctggaga
     5461 tgccaccact gcttacgcta actcggtttt taacatttgt caagccgtta ccgccaacgt
     5521 aaacgctttg ctgtctacgg atggtaacaa gatcgctgat aagtatgtgc gcaacttgca
     5581 acatagactt tatgaatgtt tgtatagaaa tcgagatgtc gatactgact tcgttaacga
     5641 attttacgcg tacttaagaa agcacttctc catgatgatt ttgtcagacg acgctgttgt
     5701 ttgtttcaat tctacttacg ctagccaagg tttggtcgct tcaatcaaaa acttcaagtc
     5761 tgttttatac taccaaaaca atgtttttat gtccgaggca aaatgctgga cggaaacaga
     5821 tttgactaag ggtccacatg aattttgtag tcaacacaca atgttagtga aacaagggga
     5881 cgactacgtt tacctaccct acccagatcc aagcagaatt ttgggtgccg gttgtttcgt
     5941 ggacgacata gtcaagaccg acggtacctt gatgatcgaa cggtttgtct ctctggccat
     6001 agacgcttac ccattgacta agcaccctaa ccaagaatat gctgatgttt ttcatttata
     6061 tcttcaatac attagaaaat tacatgacga attgaccggt cacatgttag atatgtactc
     6121 tgtgatgttg acgaacgata atacgtcacg ttactgggaa cccgaatttt acgaagcgat
     6181 gtacactcca cacacagtat tgcagtaaac aggccccttt tcctttgtcg atatcatgta
     6241 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     6301 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     6361 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     6421 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     6481 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     6541 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     6601 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     6661 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     6721 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     6781 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     6841 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     6901 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     6961 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     7021 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     7081 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     7141 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     7201 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     7261 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     7321 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     7381 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     7441 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     7501 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     7561 ttttca
//
