LOCUS       sequence-297805-        5130 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5130)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5130)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046700
FEATURES             Location/Qualifiers
     source          1..5130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(885..903)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1254..2057
                     /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      2140..3122
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3200..3229
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3251..3838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3850..3877)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3901..4092)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4093..4884,4885..4953))
                     /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     4716..4735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4954..5056)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5057..5088)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 aaattcttgg tttttcttgg tatcataact acagtagccg ctttccacca ggaatgttcc
      541 ttacaaagct gcacgcaaca tcaaccgtac gtcgttgatg acccatgtcc aatccacttc
      601 tattcgaagt ggtacattag agttggagct aggaagtctg cacctttgat tgagctgtgt
      661 gtcgatgaag ctggttcaaa gtctccaatt caatacatcg acattggtaa ttatactgtc
      721 tcctgtttgc cctttaccat taactgccaa gaaccaaaat taggcagttt ggttgtccga
      781 tgttctttct acgaagactt tttggaatat catgatgtgc gtgttgtcct agatttcatc
      841 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
      901 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
      961 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     1021 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     1081 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     1141 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     1201 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     1261 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     1321 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     1381 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     1441 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     1501 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     1561 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     1621 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     1681 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     1741 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     1801 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     1861 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     1921 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     1981 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     2041 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     2101 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     2161 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2221 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     2281 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     2341 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     2401 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     2461 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     2521 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     2581 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     2641 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     2701 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     2761 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     2821 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     2881 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     2941 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     3001 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     3061 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     3121 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     3181 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     3241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     3361 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3421 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3481 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3541 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3601 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3661 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3721 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3781 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3841 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     3901 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     3961 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     4021 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     4081 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4141 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     4201 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4261 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     4321 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4381 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4441 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4501 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     4561 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4621 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4681 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4741 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     4801 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4861 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4921 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     4981 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     5041 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     5101 ggtcaggtat gatttaaatg gtcagttaga
//
