LOCUS       Exported                6605 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4134448
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6605)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6605)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6605
                     /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             1027..1092
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1145..1266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1156..1175)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1210..1229
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1381..1427
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1457..1593
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1638..1709
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1935..2523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2015..2034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3023..4232
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4069..4088
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4236..4835
                     /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(4236..4255)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4732..4752
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4862..4910
                     /note="synthetic polyadenylation signal"
     CDS             5176..6036
                     /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(5394..5413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6049..6185
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6337..6408
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6413..6459
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6477..6525
                     /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 tcgatggatt ataaggacca cgatggagac tacaaagatc atgacattga ttacaaggac
     1081 gacgatgaca aataagctag cttgactgac tgagatacag cgtaccttca gctcacagac
     1141 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1201 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1261 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1321 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
     1381 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
     1441 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     1501 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     1561 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
     1621 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
     1681 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
     1741 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     1801 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     1861 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2101 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2341 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     2761 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     2821 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     2881 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     2941 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     3001 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     3061 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
     3121 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     3181 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     3241 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     3301 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     3361 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     3421 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     3481 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     3541 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     3601 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     3661 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     3721 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     3781 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     3841 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     3901 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     3961 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     4021 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     4081 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     4141 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
     4201 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
     4261 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4321 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4381 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4441 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4501 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4561 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4621 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4681 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4741 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4801 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
     4861 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     4921 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     4981 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     5041 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     5101 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     5161 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5221 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5281 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5341 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     5401 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5461 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     5521 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5581 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5641 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5701 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     5761 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     5821 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5881 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5941 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6001 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     6061 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     6121 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     6181 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     6241 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     6301 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     6361 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     6421 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     6481 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     6541 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     6601 ttccg
//
