LOCUS       Exported                6179 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252838
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6179)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6179
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             41..61
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             62..88
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    141..262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(152..171)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     206..225
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      377..423
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      453..589
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    634..705
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(931..1519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1011..1030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2019..3228
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3065..3084
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3232..3831
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3232..3251)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3728..3748
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3858..3906
                     /note="synthetic polyadenylation signal"
     CDS             4172..5032
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4390..4409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5045..5181
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5333..5404
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5409..5455
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5473..5521
                     /note="synthetic polyadenylation signal"
     enhancer        5602..5905
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5907..6109
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6060..6080
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6178..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gtacccctat gacgtaccag attatgcgta agctagcttg actgactgag atacagcgta
      121 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      181 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      241 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      301 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
      361 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      421 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
      481 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
      541 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
      601 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
      661 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
      721 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
      781 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
      841 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
      901 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      961 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1021 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1081 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1141 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1201 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1261 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1321 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1381 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1441 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1501 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1561 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1621 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1681 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1741 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     1801 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     1861 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     1921 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     1981 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     2041 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     2101 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     2161 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     2221 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     2281 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     2341 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     2401 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     2461 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     2521 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     2581 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     2641 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     2701 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     2761 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     2821 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     2881 tataatgcag ggtggggcca ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg
     2941 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     3001 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     3061 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     3121 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     3181 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag
     3241 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3301 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3361 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3421 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3481 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3541 gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     3601 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3661 gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc
     3721 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     3781 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg
     3841 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     3901 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     3961 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     4021 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     4081 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     4141 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4261 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     4381 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4441 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4561 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     4741 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     5041 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     5101 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     5161 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     5221 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     5281 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     5341 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     5401 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     5461 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     5521 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     5581 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5641 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5701 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5761 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     5821 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     5881 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     5941 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6001 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6061 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     6121 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
