LOCUS       Exported                6176 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252853
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6176)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6176
                     /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..85
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    138..259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(149..168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     203..222
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      374..420
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      450..586
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    631..702
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(928..1516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1008..1027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2016..3225
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3062..3081
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3229..3828
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3229..3248)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3725..3745
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3855..3903
                     /note="synthetic polyadenylation signal"
     CDS             4169..5029
                     /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(4387..4406)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5042..5178
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5330..5401
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5406..5452
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5470..5518
                     /note="synthetic polyadenylation signal"
     enhancer        5599..5902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5904..6106
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6057..6077
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgcac catcaccatc accattaagc tagcttgact gactgagata cagcgtacct
      121 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      181 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      241 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
      301 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
      361 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      421 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
      481 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
      541 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
      601 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
      661 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
      721 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
      781 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
      841 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
      901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1741 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     1801 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     1861 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     1921 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     1981 cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc
     2041 tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag
     2101 cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc
     2161 ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca
     2221 ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct
     2281 gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg
     2341 gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt
     2401 gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc
     2461 cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag
     2521 caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt
     2581 cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat
     2641 ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg
     2701 ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     2761 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     2821 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     2881 aatgcagggt ggggccacca gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     2941 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     3001 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     3061 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     3121 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt
     3181 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat gaccgagtac
     3241 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     3301 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     3361 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     3421 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     3481 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     3541 gcgcagcaac agatggaggg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     3601 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     3661 ctccccggag tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     3721 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     3781 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc tagtatgtaa
     3841 gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt
     3901 gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac
     3961 gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc
     4021 aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag
     4081 atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca
     4141 agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat
     4201 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4261 aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag
     4321 cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa
     4381 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4441 ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct
     4501 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4561 tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca
     4621 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4681 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact
     4741 attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc
     4801 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4861 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4921 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4981 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg
     5041 tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     5101 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     5161 ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct
     5221 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
     5281 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
     5341 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
     5401 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
     5461 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
     5521 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
     5581 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     5641 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     5701 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     5761 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     5821 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     5881 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
     5941 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     6001 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     6061 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     6121 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
