LOCUS       Exported                5919 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046267
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5919)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5919)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5919
                     /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             514..531
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1674..1692)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2043..2846
                     /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      2929..3911
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3989..4018
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4040..4627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4639..4666
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4690..4881)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4882..5742)
                     /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     5505..5524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5743..5845)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5846..5877
                     /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 gctgatgcac aatcgttttt aaacgggttt gcgctcgtgc cgagaggttc taaaatcgag
      541 gaaggtaagc tagtcatttg gataaatgga gataagggtt acaacggttt ggcagaagta
      601 ggcaagaaat ttgaaaagga cacaggtatc aaggtgacgg ttgaacatcc agacaagtta
      661 gaagaaaaat tccctcaagt tgctgctact ggtgatggtc ccgacattat tttctgggcc
      721 cacgatcgtt ttggtggata tgctcagtca ggtttgctgg ctgagatcac cccagacaag
      781 gccttccaag ataaattgta cccattcact tgggatgctg tcagatacaa cggtaagctt
      841 atagcatatc caattgctgt tgaagcctta tcgttgatct acaataagga cttgttacct
      901 aacccaccaa agacatggga agaaattcca gctttggata aagagctaaa ggccaaggga
      961 aaaagcgctt tgatgtttaa cttacaagaa ccttatttca cctggccatt aatcgctgct
     1021 gacggtggtt acgcatttaa atacgaaaac ggtaagtatg acattaagga tgtcggcgtt
     1081 gataatgctg gtgcaaaggc tggtttgacc ttcttggttg atttaataaa gaataaacac
     1141 atgaacgctg acactgacta ctctatcgcc gaagccgctt tcaacaaggg tgaaactgct
     1201 atgaccataa atggtccatg ggcctggagt aacattgata cttctaaagt aaactatggt
     1261 gtcactgttt tgcctacatt caaaggccaa ccatccaagc cctttgtcgg tgttctatcc
     1321 gctggcatca atgctgcttc tccaaacaag gagctagcta aagaattttt ggaaaattac
     1381 ttgttaactg acgaaggttt ggaggccgtt aacaaggaca agccattggg tgctgtcgct
     1441 ttgaaatctt acgaagaaga attggctaag gatcccagaa ttgccgctac catggagaac
     1501 gcccaaaagg gtgaaattat gccaaacatc ccacaaatgt cagccttctg gtacgcagtt
     1561 aggactgccg tgattaatgc tgcttccggt agacaaaccg tcgacgaagc attaaaagat
     1621 gctcaaacgt aaacaggccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc
     1681 ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     1741 gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta
     1801 tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta gtgcactgca
     1861 gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     1921 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     1981 attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa
     2041 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     2101 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     2161 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     2221 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     2281 tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata
     2341 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     2401 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     2461 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     2521 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     2581 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     2641 tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg
     2701 tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     2761 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     2821 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     2881 actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa cgaagcatct
     2941 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     3001 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     3061 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     3121 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     3181 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     3241 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     3301 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     3361 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     3421 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     3481 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     3541 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     3601 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     3661 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     3721 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     3781 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     3841 ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     3901 atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa gcttgactcc
     3961 agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt
     4021 agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4081 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4141 ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4501 aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata gtcctgtcgg
     4561 gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4621 atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacattt
     4681 gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     4741 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     4801 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     4861 ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4921 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatgcg
     4981 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5041 tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa gtgatcctgc
     5101 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5161 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5221 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5281 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg ccagaagtaa
     5341 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5401 gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5461 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5521 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta aactctcaag
     5581 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5641 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5701 aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt ccttagctcc
     5761 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     5821 gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct tttgactttc
     5881 tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
