LOCUS       Exported                4932 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046448
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4932)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4932)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4932
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..642
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(687..705)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1056..1859
                     /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      1942..2924
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3002..3031
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3053..3640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3652..3679
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3703..3894)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3895..4755)
                     /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     4518..4537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4756..4858)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      4859..4890
                     /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 ttctcaccat catcaccatc attaaacagg ccccttttcc
      661 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
      721 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
      781 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
      841 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
      901 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
      961 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     1021 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     1081 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     1141 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     1201 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     1261 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     1321 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     1381 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     1441 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     1501 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     1561 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     1621 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     1681 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     1741 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     1801 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     1861 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     1921 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     1981 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2041 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     2101 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     2161 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     2221 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     2281 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     2341 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     2401 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     2461 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     2521 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     2581 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     2641 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     2701 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     2761 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     2821 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     2881 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     2941 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     3001 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     3061 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3121 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     3181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3541 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     3601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     3661 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     3721 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     3781 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3841 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     3901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3961 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     4021 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4081 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     4141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4261 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4321 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     4381 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     4441 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4501 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4561 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     4621 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4681 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4741 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     4801 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     4861 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     4921 tggtcagtta ga
//
