LOCUS       Exported                6615 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252948
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6615)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6615)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6615
                     /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..79
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    148..269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(159..178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     213..232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      384..430
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      460..596
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    641..712
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(938..1526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1018..1037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2026..3235
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3072..3091
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3239..4267
                     /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    4294..4342
                     /note="synthetic polyadenylation signal"
     CDS             4608..5468
                     /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(4826..4845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5481..5617
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5769..5840
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5845..5891
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5909..5957
                     /note="synthetic polyadenylation signal"
     enhancer        6038..6341
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6343..6545
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6496..6516
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6614..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gcaccatcac catcaccatt aaggatccgc aggcctctgc tagcttgact gactgagata
      121 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
      181 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      241 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      301 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      361 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
      421 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
      481 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
      541 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
      601 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
      661 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
      721 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
      781 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
      841 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
      901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1381 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1441 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1501 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1561 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1621 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1681 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1741 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     1801 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     1861 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     1921 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     1981 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     2041 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     2101 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     2161 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     2221 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     2281 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     2341 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     2401 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     2461 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     2521 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     2581 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     2641 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     2701 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     2761 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     2821 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     2881 gttggcttat aatgcagggt ggggccacca gccggtaggt gtgcggtagg cttttctccg
     2941 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     3001 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     3061 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     3121 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     3181 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     3241 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3301 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3361 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3421 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat
     3481 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3541 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga
     3601 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3661 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     3721 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     3781 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     3841 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     3901 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     3961 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     4021 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     4081 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4141 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg
     4201 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4261 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     4321 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     4381 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4441 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4501 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4561 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4621 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4681 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     4741 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     4801 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4861 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     4921 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4981 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     5041 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5101 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5161 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     5221 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5281 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5341 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5401 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5461 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5521 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5581 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     5641 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     5701 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     5761 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     5821 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     5881 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     5941 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     6001 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6061 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6121 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6181 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6241 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6301 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     6361 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6421 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6481 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6541 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     6601 cccgccagcg gccgc
//
