LOCUS       sequence-300411-        6199 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 ORF7a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6199)
  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-m4046365
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(312..1115)
                     /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"
     primer_bind     1466..1484
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2627..2644)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(3021..3434)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      3480..3511
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3512..3614
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3615..3683,3684..4475)
                     /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     complement(3833..3852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4476..4667
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4691..4718
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4730..5317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5339..5368)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(5446..6199,1..229)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
       61 gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt
      121 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
      181 ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgtta ctcgggtaat
      241 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
      301 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
      361 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
      421 ataataatgt cagatcctgt agaaaccaca tcatccacgg ttctatactg ttgacccaat
      481 gcatctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
      541 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
      601 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
      661 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
      721 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
      781 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
      841 ctgtcctcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
      901 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
      961 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     1021 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     1081 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcacgtt
     1141 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     1201 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     1261 attaccgaat caaaaaaatt tcaaggaaac cgtgtactgc agtgcactag tggttcggcg
     1321 gccgctttgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct taatactaac
     1381 ataactatta aaaaaaataa atagggacct agacttcagg ttgtctaact ccttcctttt
     1441 cggttagagc ggatgtggga ggagggcgtg aatgtaagcg tgacataact aattacatga
     1501 tatcgacaaa ggaaaagggg cctgtttacg tttgagcatc ttttaatgct tcgtcgacgg
     1561 tttgtctacc ggaagcagca ttaatcacgg cagtcctaac tgcgtaccag aaggctgaca
     1621 tttgtgggat gtttggcata atttcaccct tttgggcgtt ctccatggta gcggcaattc
     1681 tgggatcctt agccaattct tcttcgtaag atttcaaagc gacagcaccc aatggcttgt
     1741 ccttgttaac ggcctccaaa ccttcgtcag ttaacaagta attttccaaa aattctttag
     1801 ctagctcctt gtttggagaa gcagcattga tgccagcgga tagaacaccg acaaagggct
     1861 tggatggttg gcctttgaat gtaggcaaaa cagtgacacc atagtttact ttagaagtat
     1921 caatgttact ccaggcccat ggaccattta tggtcatagc agtttcaccc ttgttgaaag
     1981 cggcttcggc gatagagtag tcagtgtcag cgttcatgtg tttattcttt attaaatcaa
     2041 ccaagaaggt caaaccagcc tttgcaccag cattatcaac gccgacatcc ttaatgtcat
     2101 acttaccgtt ttcgtattta aatgcgtaac caccgtcagc agcgattaat ggccaggtga
     2161 aataaggttc ttgtaagtta aacatcaaag cgctttttcc cttggccttt agctctttat
     2221 ccaaagctgg aatttcttcc catgtctttg gtgggttagg taacaagtcc ttattgtaga
     2281 tcaacgataa ggcttcaaca gcaattggat atgctataag cttaccgttg tatctgacag
     2341 catcccaagt gaatgggtac aatttatctt ggaaggcctt gtctggggtg atctcagcca
     2401 gcaaacctga ctgagcatat ccaccaaaac gatcgtgggc ccagaaaata atgtcgggac
     2461 catcaccagt agcagcaact tgagggaatt tttcttctaa cttgtctgga tgttcaaccg
     2521 tcaccttgat acctgtgtcc ttttcaaatt tcttgcctac ttctgccaaa ccgttgtaac
     2581 ccttatctcc atttatccaa atgactagct taccttcctc gattttagaa cctctcggca
     2641 cgagttcagt ctttcgcttc agtgtgaaac acaaggtaat gaaaacgata gcggcgacaa
     2701 ttagaaaaat tggggagtac aactcttgaa cttcttcttg acggataaat aactttgggc
     2761 ttacggatct cgctctcaat tgataaacgt gtttcacacc gtcgggacaa gcgaaagcga
     2821 attgagtact aaaacaggtt aaggcgaact tgttatcagc caatggatga aatggcgaat
     2881 tgccttcgta agttccagat gaacaaggtt ccttcaacaa gacagttgtc cccctaacgc
     2941 attcctggta gtgataaagc tcacacgtag ccagggttat caatgctaag aataggataa
     3001 ttttcatttt gtaattaaaa cttagattag attgctatgc tttctttcta atgagcaaga
     3061 agtaaaaaaa gttgtaatag aacaagaaaa atgaaactga aacttgagaa attgaagacc
     3121 gtttattaac ttaaatatca atgggaggtc atcgaaagag aaaaaaatca aaaaaaaaaa
     3181 ttttcaagaa aaagaaacgt gataaaaatt tttattgcct ttttcgacga agaaaaagaa
     3241 acgaggcggt ctcttttttc ttttccaaac ctttagtacg ggtaattaac gacaccctag
     3301 aggaagaaag aggggaaatt tagtatgctg tgcttgggtg ttttgaagtg gtacggcgat
     3361 gcgcggagtc cgagaaaatc tggaagagta aaaaaggagt agaaacattt tgaagctatg
     3421 gtgtgtgggg cggccgctct aactgaccat ttaaatcata cctgacctcc atagcagaaa
     3481 gtcaaaagcc tccgaccgga ggcttttgac ttgatcggca cgtaagaggt tccaactttc
     3541 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     3601 ctaaggaagc taaaatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3781 agagtttacg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatctcacg gatggcatga
     3961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4021 ttctggcaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4081 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4261 gatcacttct gcgctcggcc ctcccggctg gctggtttat tgctgataaa tctggagccg
     4321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgca
     4381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4441 ctgagatagg tgcctcactg attaagcatt ggtaagtgac caaacaggaa aaaaccgccc
     4501 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     4561 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     4621 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgatga gggcccaaat
     4681 gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt gctggcgttt ttccataggc
     4741 tccgcccccc tgacgagcat cacaaaaatc gatgctcaag tcagaggtgg cgaaacccga
     4801 caggactata aagataccag gcgtttaccc ctggaagctc cctcgtgcgc tctcctgttc
     4861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5161 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc tcggaaaaag
     5221 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5281 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga ttttctaccg
     5341 aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gcagtccagt tacgctggag
     5401 tcaagcttgt aacacgcggc cgctcgggaa gttcctattc tctagctaga aagtatagga
     5461 acttcagagc gcttttgaaa accaaaagcg ctctgatgac gcactttcaa aaaaccaaaa
     5521 acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct
     5581 caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct acccatccac
     5641 ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt ttatctctag
     5701 tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa tcgaaaacaa
     5761 tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa gaaaccgttc
     5821 ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac aaagtatgcg
     5881 caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa atgcacccgc
     5941 agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta tggaagagaa
     6001 aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa aaagttatca
     6061 agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg ctttgttaga
     6121 aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga ttctttgttg
     6181 gtaaaatagc gctctcgcg
//
