LOCUS       Exported                6609 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046305
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6609)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6609)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6609
                     /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             2278..2295
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2296..2319
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2364..2382)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2733..3536
                     /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      3619..4601
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4679..4708
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4730..5317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      5329..5356
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5380..5571)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5572..6432)
                     /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     6195..6214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6433..6535)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6536..6567
                     /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 gttggggctt gtgttctttg caattcacag acttcattaa gatgtggtgc ttgcatacgt
      541 agaccattct tatgttgtaa atgctgttac gaccatgtca tatcaacatc acataaatta
      601 gtcttgtctg ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac agatgtgact
      661 caactttact taggaggtat gagctattat tgtaaatcac ataaaccacc cattagtttt
      721 ccattgtgtg ctaatggaca agtttttggt ttatataaaa atacatgtgt tggtagcgat
      781 aatgttactg actttaatgc aattgcaaca tgtgactgga caaatgctgg tgattacatt
      841 ttagctaaca cctgtactga aagactcaag ctttttgcag cagaaacgct caaagctact
      901 gaggagacat ttaaactgtc ttatggtatt gctactgtac gtgaagtgct gtctgacaga
      961 gaattacatc tttcatggga agttggtaaa cctagaccac cacttaaccg aaattatgtc
     1021 tttactggtt atcgtgtaac taaaaacagt aaagtacaaa taggagagta cacctttgaa
     1081 aaaggtgact atggtgatgc tgttgtttac cgaggtacaa caacttacaa attaaatgtt
     1141 ggtgattatt ttgtgctgac atcacataca gtaatgccat taagtgcacc tacactagtg
     1201 ccacaagagc actatgttag aattactggc ttatacccaa cactcaatat ctcagatgag
     1261 ttttctagca atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc tacactccag
     1321 ggaccacctg gtactggtaa gagtcatttt gctattggcc tagctctcta ctacccttct
     1381 gctcgcatag tgtatacagc ttgctctcat gccgctgttg atgcactatg tgagaaggca
     1441 ttaaaatatt tgcctataga taaatgtagt agaattatac ctgcacgtgc tcgtgtagag
     1501 tgttttgata aattcaaagt gaattcaaca ttagaacagt atgtcttttg tactgtaaat
     1561 gcattgcctg agacgacagc agatatagtt gtctttgatg aaatttcaat ggccacaaat
     1621 tatgatttga gtgttgtcaa tgccagatta cgtgctaagc actatgtgta cattggcgac
     1681 cctgctcaat tacctgcacc acgcacattg ctaactaagg gcacactaga accagaatat
     1741 ttcaattcag tgtgtagact tatgaaaact ataggtccag acatgttcct cggaacttgt
     1801 cggcgttgtc ctgctgaaat tgttgacact gtgagtgctt tggtttatga taataagctt
     1861 aaagcacata aagacaaatc agctcaatgc tttaaaatgt tttataaggg tgttatcacg
     1921 catgatgttt catctgcaat taacaggcca caaataggcg tggtaagaga attccttaca
     1981 cgtaaccctg cttggagaaa agctgtcttt atttcacctt ataattcaca gaatgctgta
     2041 gcctcaaaga ttttgggact accaactcaa actgttgatt catcacaggg ctcagaatat
     2101 gactatgtca tattcactca aaccactgaa acagctcact cttgtaatgt aaacagattt
     2161 aatgttgcta ttaccagagc aaaagtaggc atactttgca taatgtctga tagagacctt
     2221 tatgacaagt tgcaatttac aagtcttgaa attccacgta ggaatgtggc aactttactc
     2281 gtgccgagag gttcttggag ccatccgcag tttgaaaaat aaacaggccc cttttccttt
     2341 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     2401 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     2461 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     2521 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     2581 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     2641 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     2701 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     2761 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     2821 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     2881 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     2941 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     3001 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     3061 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     3121 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     3181 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     3241 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     3301 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     3361 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     3421 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     3481 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     3541 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     3601 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     3661 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3721 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     3781 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     3841 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     3901 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     3961 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     4021 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     4081 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     4141 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     4201 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     4261 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     4321 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     4381 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     4441 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     4501 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     4561 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     4621 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     4681 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4801 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     4861 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5101 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     5221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     5281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     5341 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     5401 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5461 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5521 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5641 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5761 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6001 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     6061 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6241 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6421 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     6481 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     6541 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     6601 tcagttaga
//
