LOCUS       Exported                6129 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046304
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6129
                     /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             724..741
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1884..1902)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2253..3056
                     /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      3139..4121
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4199..4228
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4250..4837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4849..4876
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4900..5091)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5092..5952)
                     /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     5715..5734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5953..6055)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6056..6087
                     /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 aaaatgtcag atgtaaagtg cacatcagta gtcttactct cagttttgca acaactcaga
      541 gtagaatcat catctaaatt gtgggctcaa tgtgtccagt tacacaatga cattctctta
      601 gctaaagata ctactgaagc ctttgaaaaa atggtttcac tactttctgt tttgctttcc
      661 atgcagggtg ctgtagacat aaacaagctt tgtgaagaaa tgctggacaa cagggcaacc
      721 ttactcgtgc cgagaggttc taaaatcgag gaaggtaagc tagtcatttg gataaatgga
      781 gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga cacaggtatc
      841 aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt tgctgctact
      901 ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata tgctcagtca
      961 ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta cccattcact
     1021 tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt tgaagcctta
     1081 tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga agaaattcca
     1141 gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa cttacaagaa
     1201 ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa atacgaaaac
     1261 ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc tggtttgacc
     1321 ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta ctctatcgcc
     1381 gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg ggcctggagt
     1441 aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt caaaggccaa
     1501 ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc tccaaacaag
     1561 gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt ggaggccgtt
     1621 aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga attggctaag
     1681 gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat gccaaacatc
     1741 ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc tgcttccggt
     1801 agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc cttttccttt
     1861 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     1921 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     1981 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     2041 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     2101 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     2161 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     2221 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     2281 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     2341 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     2401 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     2461 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     2521 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     2581 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     2641 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     2701 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     2761 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     2821 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     2881 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     2941 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     3001 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     3061 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     3121 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     3181 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3241 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     3301 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     3361 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     3421 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     3481 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     3541 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     3601 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     3661 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     3721 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     3781 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     3841 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     3901 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     3961 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     4021 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     4081 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     4141 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     4201 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     4261 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4321 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     4381 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4501 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4561 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4621 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4681 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     4741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     4801 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     4861 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     4921 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     4981 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5041 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     5101 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5161 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     5221 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5281 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     5341 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5401 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5461 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5521 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     5581 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     5641 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5701 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5761 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5821 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5881 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5941 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     6001 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     6061 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     6121 tcagttaga
//
