LOCUS       Exported                6630 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252960
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6630)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6630)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6630
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             47..67
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             68..94
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    163..284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(174..193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     228..247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      399..445
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      475..611
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    656..727
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(953..1541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1033..1052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2041..3250
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3087..3106
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3254..4282
                     /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    4309..4357
                     /note="synthetic polyadenylation signal"
     CDS             4623..5483
                     /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(4841..4860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5496..5632
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5784..5855
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5860..5906
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5924..5972
                     /note="synthetic polyadenylation signal"
     enhancer        6053..6356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6358..6560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6511..6531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgtac ccctatgacg taccagatta tgcgtaagga tccgcaggcc tctgctagct
      121 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
      181 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
      241 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
      301 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
      361 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
      421 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
      481 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
      541 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
      601 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
      661 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
      721 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
      781 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
      841 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
      901 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      961 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1021 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1081 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1141 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1201 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1261 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1321 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1381 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1441 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1501 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1561 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1621 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1681 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1741 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     1801 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     1861 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     1921 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     1981 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     2041 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg
     2101 ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc
     2161 ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg
     2221 acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga
     2281 aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg
     2341 attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc
     2401 gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg
     2461 gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc
     2521 aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc
     2581 agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag
     2641 gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc
     2701 gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga
     2761 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     2821 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     2881 gacgtcaccc gttctgttgg cttataatgc agggtggggc caccagccgg taggtgtgcg
     2941 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     3001 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     3061 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     3121 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
     3181 atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt
     3241 tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     3301 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     3361 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     3421 tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc tcccgattcc
     3481 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     3541 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     3601 cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg ggttcggccc
     3661 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     3721 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     3781 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     3841 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     3901 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     3961 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4021 gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg accaactcta
     4081 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     4141 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgccgc
     4201 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     4261 tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa taaaatcgct
     4321 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg
     4381 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     4441 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     4501 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     4561 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     4621 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4681 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4741 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4801 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4861 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4921 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4981 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5041 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5101 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5161 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     5221 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     5281 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5341 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5401 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5461 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     5521 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     5581 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     5641 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     5701 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     5761 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     5821 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     5881 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     5941 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     6001 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     6061 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     6121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     6181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     6241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     6301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     6361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     6421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     6481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     6541 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     6601 catccactcg acacacccgc cagcggccgc
//
