LOCUS       Exported                6395 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133301
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6395)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6395)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6395
                     /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"
     CDS             835..855
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             856..882
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    935..1056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(946..965)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1000..1019
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1171..1217
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1247..1383
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1428..1499
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1725..2313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1805..1824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2813..4022
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3859..3878
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4026..4625
                     /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(4026..4045)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4522..4542
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4652..4700
                     /note="synthetic polyadenylation signal"
     CDS             4966..5826
                     /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(5184..5203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5839..5975
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6127..6198
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6203..6249
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6267..6315
                     /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 ccatgagcaa aatgtctgac
      601 gtcaagtgta catccgtagt tctgttatca gtgcttcaac agctgcgggt ggagagttcg
      661 tcaaagctct gggcccagtg cgtgcagctg cacaatgata tattgctcgc gaaagatact
      721 acggaagctt tcgagaagat ggtcagcttg ctatctgttc tactgagcat gcaaggagca
      781 gtagacatca acaaactttg cgaagagatg ttagacaacc gagccaccct gcaagaaaac
      841 ctttactttc agtcgtaccc ctatgacgta ccagattatg cgtaagctag cttgactgac
      901 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
      961 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1021 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1081 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1141 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
     1201 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
     1261 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     1321 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     1381 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
     1441 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
     1501 tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg
     1561 agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt
     1621 ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc
     1681 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1741 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1801 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1861 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1921 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1981 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2041 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2101 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2161 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2221 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2281 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2341 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2401 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2461 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2521 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
     2581 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
     2641 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
     2701 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
     2761 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg
     2821 cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca
     2881 gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
     2941 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
     3001 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
     3061 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
     3121 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
     3181 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
     3241 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
     3301 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
     3361 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
     3421 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc
     3481 gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt
     3541 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     3601 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     3661 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
     3721 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     3781 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     3841 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     3901 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     3961 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     4021 acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4081 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     4141 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4201 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4261 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4321 gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg caccggccca
     4381 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4441 tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg gtgcccgcct
     4501 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     4561 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     4621 cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat
     4681 gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc
     4741 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     4801 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     4861 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     4921 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     4981 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5041 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     5101 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     5161 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5221 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     5281 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5341 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     5401 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5461 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5521 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     5581 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5641 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5701 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5761 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5821 tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     5881 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     5941 catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact
     6001 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     6061 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     6121 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     6181 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     6241 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
     6301 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
     6361 gggtcattag ttcatagccc atatatggag ttccg
//
