LOCUS       Exported                5109 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046668
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5109)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5109)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5109
                     /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"
     primer_bind     complement(864..882)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1233..2036
                     /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      2119..3101
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3179..3208
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3230..3817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3829..3856
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3880..4071)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4072..4932)
                     /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     4695..4714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4933..5035)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5036..5067
                     /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 aacaatgagc tttctcctgt tgctttacgg cagatgtcct gcgcagctgg aacaacgcaa
      541 actgcctgta ctgacgataa cgctctagct tactataaca ccactaaagg tggtagattc
      601 gtcttggcct tgttgtcaga cctgcaagat ttgaagtggg cgagatttcc aaagagcgat
      661 ggcactggta ccatatacac cgaattagaa ccaccctgtc gtttcgtgac cgacactcca
      721 aaaggtccga aggttaagta tttgtacttt attaagggtt taaataactt gaatcgaggt
      781 atggttctgg ggtctttggc tgctacagtc agactccaat aaacaggccc cttttccttt
      841 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
      901 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
      961 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     1021 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     1081 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     1141 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     1201 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     1261 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     1321 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     1381 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     1441 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     1501 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     1561 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     1621 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     1681 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     1741 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     1801 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     1861 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     1921 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     1981 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     2041 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     2101 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     2161 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2221 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     2281 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2341 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     2401 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     2461 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     2521 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     2581 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     2641 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     2701 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     2761 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     2821 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     2881 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     2941 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     3001 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     3061 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     3121 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     3181 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     3241 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3301 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     3361 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3421 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3481 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3541 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3601 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3661 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     3721 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     3781 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     3841 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     3901 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     3961 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     4021 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     4081 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4141 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     4201 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4261 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     4321 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4381 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4441 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4501 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     4561 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     4621 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4681 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4741 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4801 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4861 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4921 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     4981 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     5041 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     5101 tcagttaga
//
