LOCUS       Exported                6624 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252940
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6624)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6624)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6624
                     /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    157..278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(168..187)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     222..241
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      393..439
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      469..605
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    650..721
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(947..1535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1027..1046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2035..3244
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3081..3100
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3248..4276
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4303..4351
                     /note="synthetic polyadenylation signal"
     CDS             4617..5477
                     /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(4835..4854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5490..5626
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5778..5849
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5854..5900
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5918..5966
                     /note="synthetic polyadenylation signal"
     enhancer        6047..6350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6352..6554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6505..6525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6623..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 aggatccgca ggcctctgct agcttgactg
      121 actgagatac agcgtacctt cagctcacag acatgataag atacattgat gagtttggac
      181 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      241 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      301 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      361 aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg
      421 ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag
      481 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
      541 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
      601 cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa
      661 tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct
      721 aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg
      781 cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg
      841 gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag
      901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1381 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1501 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1621 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1681 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1741 cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct
     1801 cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac
     1861 tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca
     1921 ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca
     1981 tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc
     2041 cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt
     2101 cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg
     2161 ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga
     2221 cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta
     2281 gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata
     2341 taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt
     2401 cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg
     2461 gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc
     2521 tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa
     2581 atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt
     2641 gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat
     2701 gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa
     2761 gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg
     2821 tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc
     2881 acccgttctg ttggcttata atgcagggtg gggccaccag ccggtaggtg tgcggtaggc
     2941 ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc
     3001 gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg
     3061 tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag
     3121 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa
     3181 ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt
     3241 agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     3301 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     3361 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     3421 ctacaaagat cgttatgttt atcggcactt tgcatccgcc gcgctcccga ttccggaagt
     3481 gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     3541 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3601 ggcaatggat gccatcgctg ccgccgatct tagccagacg agcgggttcg gcccattcgg
     3661 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3721 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     3781 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     3841 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     3901 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     3961 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     4021 aggatcgccc cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     4081 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     4141 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ccgccgtctg
     4201 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     4261 gagggcaaag gaataagcta gtatgtaagc ctagtcttag ataataaaat cgctatccat
     4321 cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga
     4381 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     4441 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     4501 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     4561 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     4621 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4681 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     4741 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4801 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4861 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4921 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4981 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     5041 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5101 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5161 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     5221 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5281 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5341 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5401 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5461 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     5521 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     5581 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     5641 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     5701 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     5761 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     5821 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     5881 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     5941 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     6001 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     6061 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     6121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     6181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     6241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     6301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     6361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     6421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     6481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     6541 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     6601 ctcgacacac ccgccagcgg ccgc
//
