LOCUS       Exported                6675 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252876
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6675)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6675)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6675
                     /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..139
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    208..329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(219..238)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     273..292
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      444..490
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      520..656
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    701..772
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(998..1586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1078..1097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2086..3295
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3132..3151
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3299..4327
                     /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    4354..4402
                     /note="synthetic polyadenylation signal"
     CDS             4668..5528
                     /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(4886..4905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5541..5677
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5829..5900
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5905..5951
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5969..6017
                     /note="synthetic polyadenylation signal"
     enhancer        6098..6401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6403..6605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6556..6576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6674..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gaagaggcga tggaaaaaga attttatcgc ggtgagtgca gccaacagat tcaaaaagat
      121 atcctcgtct ggtgccctct aaggatccgc aggcctctgc tagcttgact gactgagata
      181 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
      241 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      301 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      361 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      421 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
      481 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
      541 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
      601 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
      661 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
      721 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
      781 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
      841 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
      901 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
      961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1741 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1801 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     1861 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     1921 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     1981 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     2041 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     2101 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     2161 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     2221 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     2281 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     2341 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     2401 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     2461 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     2521 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     2581 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     2641 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     2701 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     2761 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     2821 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     2881 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     2941 gttggcttat aatgcagggt ggggccacca gccggtaggt gtgcggtagg cttttctccg
     3001 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     3061 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     3121 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     3181 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     3241 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     3301 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3361 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3421 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3481 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat
     3541 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3601 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga
     3661 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3721 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     3781 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     3841 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     3901 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     3961 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     4021 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     4081 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     4141 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4201 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg
     4261 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4321 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     4381 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     4441 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4501 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4561 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4621 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     4801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     4861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     4981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5041 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     5101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5221 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     5281 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5341 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5401 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5521 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5581 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5641 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     5701 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     5761 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     5821 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     5881 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     5941 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     6001 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     6061 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6361 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     6421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6601 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     6661 cccgccagcg gccgc
//
