LOCUS       sequence-287724-        4980 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4980)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4980)
  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-m4046175
FEATURES             Location/Qualifiers
     source          1..4980
                     /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"
     CDS             607..624
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             625..690
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(735..753)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1104..1907
                     /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      1990..2972
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3050..3079
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3101..3688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3700..3727)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3751..3942)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3943..4734,4735..4803))
                     /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     4566..4585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4804..4906)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4907..4938)
                     /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 atcgagttat ccttgataga tttctatcta tgttttttgg cattcctgct ttttctcgtg
      541 ttgattatgt tgattatctt ctggttcagt ttagaattgc aagaccataa tgaaacatgc
      601 cacgccctcg tgccgagagg ttctgattat aaagatcatg atggtgatta taaagatcat
      661 gatattgatt ataaagatga tgatgataaa taaacaggcc ccttttcctt tgtcgatatc
      721 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
      781 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
      841 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
      901 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
      961 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     1021 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     1081 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     1141 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     1201 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     1261 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     1321 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     1381 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     1441 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     1501 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     1561 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     1621 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     1681 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     1741 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     1801 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     1861 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     1921 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     1981 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     2041 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     2101 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     2161 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2221 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     2281 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     2341 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     2401 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     2461 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     2521 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     2581 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     2641 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     2701 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     2761 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     2821 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     2881 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     2941 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     3001 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     3061 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     3121 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3181 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     3241 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3301 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3361 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3421 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3481 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3541 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     3601 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     3661 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     3721 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     3781 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     3841 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     3901 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     3961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4021 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     4081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     4141 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4381 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4441 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     4501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4621 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4801 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     4861 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     4921 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
