LOCUS       Exported                7145 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133386
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7145)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7145)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7145
                     /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             95..172
                     /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"
     CDS             200..220
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    685..806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(696..715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     750..769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      921..967
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      997..1133
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1178..1249
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1475..2063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1555..1574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2563..3772
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3609..3628
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3776..4804
                     /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    4831..4879
                     /note="synthetic polyadenylation signal"
     CDS             5145..6005
                     /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(5363..5382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6018..6154
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6299..6370
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6375..6421
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6439..6487
                     /note="synthetic polyadenylation signal"
     enhancer        6568..6871
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6873..7075
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7026..7046
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      7144..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gggacggcga atcccggggc tgataaatcc ctggaaaagg agatggaaga agaactttat
      121 tgcagtgtct gcggctaacc ggttcaaaaa gatcagttca agcggcgccc tagattacga
      181 cattcctacg accgcctccg agaatctcta tttccagggt gaatttggct tggcacaaca
      241 tgacgaggcc gtcgataaca agtttaataa agaacagcag aacgctttct atgagatcct
      301 tcacttacca aatctgaacg aagagcaacg caacgctttc attcagagcc tgaaagacga
      361 tccctcgcag tctgcaaatc tcctggccga agccaagaag ctgaatgacg cacaggctcc
      421 aaaagttgat aacaagttta acaaggagca gcagaatgct ttctacgaga tattgcacct
      481 tcctaatctg aatgaggaac aaagaaatgc atttatccag tccctaaaag acgatccatc
      541 acagagtgcc aacctgcttg cggaagccaa aaagctcaat gatgcccagg ccccgaaagt
      601 agacgcgaac catcaataag gatccgcagg cctctgctag cttgactgac tgagatacag
      661 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
      721 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      781 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      841 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
      901 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
      961 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     1021 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     1081 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     1141 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     1201 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     1261 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     1321 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     1381 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     1441 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1501 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1561 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1621 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1681 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1741 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1801 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1861 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1921 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1981 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2041 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2101 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2161 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2221 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2281 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     2341 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     2401 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     2461 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     2521 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     2581 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     2641 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2701 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2761 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2821 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2881 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     2941 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3001 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3061 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3121 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3181 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     3241 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     3301 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     3361 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     3421 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
     3481 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     3541 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     3601 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     3661 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     3721 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     3781 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3841 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3901 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3961 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     4021 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4081 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     4141 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     4201 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     4261 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4321 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4381 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4441 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4501 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     4561 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4621 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4681 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     4741 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4801 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     4861 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     4921 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     4981 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5041 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5101 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     5161 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5221 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     5281 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     5341 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5401 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     5461 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5521 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5581 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5641 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5701 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     5761 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5821 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5881 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5941 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6001 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     6061 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     6121 atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct
     6181 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     6241 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     6301 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     6361 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     6421 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     6481 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     6541 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     6601 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6661 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     6721 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     6781 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     6841 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     6901 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     6961 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     7021 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     7081 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     7141 gccgc
//
