LOCUS       Exported                6581 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133148
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6581)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6581)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6581
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1003..1023
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1024..1068
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1121..1242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1132..1151)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1186..1205
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1357..1403
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1433..1569
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1614..1685
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1911..2499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1991..2010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2999..4208
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4045..4064
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4212..4811
                     /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(4212..4231)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4708..4728
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4838..4886
                     /note="synthetic polyadenylation signal"
     CDS             5152..6012
                     /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(5370..5389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6025..6161
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6313..6384
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6389..6435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6453..6501
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggcagg gaatgcgaca
      601 gaggttcctg ccaactcaac ggtcttgtct ttttgtgctt tcgccgtgga cgctgcaaaa
      661 gcctacaagg attatctagc ctcgggtgga cagccaatca ctaactgcgt aaagatgctt
      721 tgtacccaca ccggcactgg ccaagcaatt acagtgaccc ccgaagctaa tatggaccag
      781 gagtccttcg gaggtgcgag ctgctgcctg tactgtcgtt gtcatataga tcacccgaat
      841 cccaaagggt tttgcgacct caagggcaaa tatgtccaga tcccaacgac atgcgccaac
      901 gatcctgtgg gattcactct gaagaacacc gtatgtaccg tttgcgggat gtggaagggg
      961 tatggctgta gttgcgacca attacgggaa ccgatgctgc aagaaaacct ttactttcag
     1021 tcgaaagaaa ccgcggcagc taagttcgag aggcagcaca tggacagtta agctagcttg
     1081 actgactgag atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt
     1141 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1201 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1261 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1321 tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta
     1381 aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc
     1441 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     1501 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     1561 tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct
     1621 ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct
     1681 ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac
     1741 ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga
     1801 tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca
     1861 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1921 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1981 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2041 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2101 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2161 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2221 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2281 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2341 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2401 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2461 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2521 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2581 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2641 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2701 tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg
     2761 ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac
     2821 gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc
     2881 gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc
     2941 tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg
     3001 cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc
     3061 acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     3121 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     3181 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     3241 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     3301 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     3361 ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc
     3421 cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa
     3481 gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag
     3541 caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt
     3601 cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc
     3661 taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg
     3721 tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg
     3781 gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga
     3841 cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt
     3901 aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac
     3961 aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt
     4021 tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg
     4081 cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat
     4141 gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt
     4201 tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4261 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4321 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4381 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4441 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4501 agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg gcgccgcacc
     4561 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4621 agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc gccggggtgc
     4681 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4741 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     4801 ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     4861 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     4921 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     4981 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     5041 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     5101 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     5161 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5221 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     5281 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     5341 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5401 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     5461 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5521 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     5581 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5641 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5701 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5761 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5821 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5881 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5941 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6001 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     6061 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     6121 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     6181 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     6241 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     6301 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     6361 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     6421 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     6481 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca
     6541 attacggggt cattagttca tagcccatat atggagttcc g
//
