LOCUS       sequence-291556-        6709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6709)
  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-m4046499
FEATURES             Location/Qualifiers
     source          1..6709
                     /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             1304..1321
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2464..2482)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2833..3636
                     /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      3719..4701
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4779..4808
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4830..5417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5429..5456)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5480..5671)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5672..6463,6464..6532))
                     /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     6295..6314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6533..6635)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6636..6667)
                     /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 atgtctgctg tcaaaagaac tattaagggt acccatcact ggttgctgct
      481 aacaatctta acttccttgt tagtgcttgt tcaaagtacc cagtggtcgt tgtttttctt
      541 cttgtacgag aacgcctttc tcccattcgc aatggggata attgctatgt cagcttttgc
      601 catgatgttc gttaagcaca aacatgcgtt tttgtgctta ttcttgttgc cttctctcgc
      661 tacggtagct tatttcaata tggtctacat gccagcttcc tgggttatgc gtatcatgac
      721 ttggttggac atggttgata ccagcctttc tggtttcaag ttgaaagatt gtgtcatgta
      781 cgcatctgcc gttgtattac taattcttat gactgctaga accgtctatg acgatggcgc
      841 gagaagggtt tggacactaa tgaacgtgtt gactttagtt tacaaggtct actatggaaa
      901 tgccttggac caagctatct ccatgtgggc tttaataatc tcagtcactt ctaactacag
      961 tggtgttgtt actacagtga tgtttttggc tagaggtatt gtcttcatgt gtgttgaata
     1021 ttgtcccatt ttctttatca ccggtaacac tttacaatgc attatgttgg tttactgttt
     1081 cttaggttac ttttgtacct gttatttcgg cctattttgt ttgttgaatc gatactttag
     1141 actcacattg ggtgtttacg attatttggt cagcactcaa gaatttagat atatgaactc
     1201 ccaaggtttg ttaccaccaa agaactctat tgatgccttc aagttgaaca tcaagctgtt
     1261 aggtgtcggt ggaaaacctt gcatcaaggt agccaccgtc caactcgtgc cgagaggttc
     1321 taaaatcgag gaaggtaagc tagtcatttg gataaatgga gataagggtt acaacggttt
     1381 ggcagaagta ggcaagaaat ttgaaaagga cacaggtatc aaggtgacgg ttgaacatcc
     1441 agacaagtta gaagaaaaat tccctcaagt tgctgctact ggtgatggtc ccgacattat
     1501 tttctgggcc cacgatcgtt ttggtggata tgctcagtca ggtttgctgg ctgagatcac
     1561 cccagacaag gccttccaag ataaattgta cccattcact tgggatgctg tcagatacaa
     1621 cggtaagctt atagcatatc caattgctgt tgaagcctta tcgttgatct acaataagga
     1681 cttgttacct aacccaccaa agacatggga agaaattcca gctttggata aagagctaaa
     1741 ggccaaggga aaaagcgctt tgatgtttaa cttacaagaa ccttatttca cctggccatt
     1801 aatcgctgct gacggtggtt acgcatttaa atacgaaaac ggtaagtatg acattaagga
     1861 tgtcggcgtt gataatgctg gtgcaaaggc tggtttgacc ttcttggttg atttaataaa
     1921 gaataaacac atgaacgctg acactgacta ctctatcgcc gaagccgctt tcaacaaggg
     1981 tgaaactgct atgaccataa atggtccatg ggcctggagt aacattgata cttctaaagt
     2041 aaactatggt gtcactgttt tgcctacatt caaaggccaa ccatccaagc cctttgtcgg
     2101 tgttctatcc gctggcatca atgctgcttc tccaaacaag gagctagcta aagaattttt
     2161 ggaaaattac ttgttaactg acgaaggttt ggaggccgtt aacaaggaca agccattggg
     2221 tgctgtcgct ttgaaatctt acgaagaaga attggctaag gatcccagaa ttgccgctac
     2281 catggagaac gcccaaaagg gtgaaattat gccaaacatc ccacaaatgt cagccttctg
     2341 gtacgcagtt aggactgccg tgattaatgc tgcttccggt agacaaaccg tcgacgaagc
     2401 attaaaagat gctcaaacgt aaacaggccc cttttccttt gtcgatatca tgtaattagt
     2461 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
     2521 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
     2581 acgttattta tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta
     2641 gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     2701 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     2761 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa
     2821 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     2881 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     2941 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     3001 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     3061 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac
     3121 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     3181 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     3241 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     3301 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     3361 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     3421 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag
     3481 tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga
     3541 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     3601 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     3661 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa
     3721 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     3781 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     3841 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
     3901 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     3961 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     4021 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     4081 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     4141 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     4201 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     4261 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     4321 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     4381 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     4441 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     4501 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     4561 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     4621 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
     4681 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
     4741 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     4801 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4861 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4921 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4981 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5041 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5101 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5161 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5221 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5281 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     5341 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     5401 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     5461 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     5521 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     5581 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     5641 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     5701 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5761 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5821 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     5881 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5941 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6001 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6061 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6121 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6181 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     6241 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6301 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     6361 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6421 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6481 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     6541 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     6601 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct
     6661 tttgactttc tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
