LOCUS       Exported                6395 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252809
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"
     polyA_signal    49..97
                     /note="synthetic polyadenylation signal"
     terminator      115..161
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    166..237
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      389..525
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(538..1398)
                     /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     1161..1180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1664..1712
                     /note="synthetic polyadenylation signal"
     CDS             complement(1739..2338)
                     /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(1822..1842)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2319..2338
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2342..3551)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2486..2505)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4051..4639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4540..4559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4865..4936
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      4981..5117
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      5147..5193
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5308..5429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5345..5364)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5399..5418
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5482..5508)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5509..5529)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5856..6058)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5885..5905)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6060..6363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccccgtaatt gattactaga tctggctaca ctcgatcagc agttagataa taaaatcgct
       61 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat
      121 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gaattgagag gtagctgaag
      181 ctgctgttat gaccactaga gggaagaaga tacctgtgga gctaatggtc caagatgtcg
      241 ggagccgcga tcgctgccaa aaaggtaagc tgacgatagc tgacgagatt cgatgcccga
      301 cgagcaaatg agcgccacgg ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc
      361 cagagtcgag tttaaacgat tcgggatctt aaggataaat ttctggtaag gaggacacgt
      421 atggaagtgg gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta
      481 taagacgcgc agcgtcgcat caggcatttt tttctgcgcc aatgcaccta gaatcggtta
      541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      781 tatcaactgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tacgcaaggt
      841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      961 tagctccttc ggtcctccaa tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1081 gactggtgaa tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1201 cattggaaag cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1261 ttcgatgtaa cccactcgcg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1381 gaaatgttga atactcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc
     1441 aaaagctagg actcgattat acgttgaccg atcagactat caatatctag gtttaaacag
     1501 atcggtacgc tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc
     1561 ccaagctgga ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac
     1621 tgagaggatc gttctcggta caccaggcgc gcctagcgtt acacacacaa aaaaccaaca
     1681 cacatccatc ttcgatggat agcgatttta ttatctaaga ctaggcttac atactagcct
     1741 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     1801 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca
     1861 ggaaggcggg caccccggcg cgctcggcgg cctccactcc ggggagcacg acggcgctgc
     1921 ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct
     1981 ccttgggccg gtgcggcgcc aggaggccct ccatctgttg ctgcgcggcc agccgggaac
     2041 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
     2101 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
     2161 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt
     2221 ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
     2281 cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgt
     2341 tgtctaacaa aaaagccaaa aacggccaga atttagcgga caatttacaa gtctaacact
     2401 gaaaattaca tattgaccca aatgattaca tttcaaaagg tgcctaaaaa acttcacaaa
     2461 acacactcgc caaccccgag cgcatagttc aaaaccggag cttcagctac ttaagaagat
     2521 aggtacataa aaccgaccaa agaaactgac gcctcactta tccctcccct caccagaggt
     2581 ccggcgcctg tcgattcagg agagcctacc ctaggcccga accctgcgtc ctgcgacgga
     2641 gaaaagccta ccgcacacct accggctggt ggccccaccc tgcattataa gccaacagaa
     2701 cgggtgacgt cacgacacga cgagggcgcg cgctcccaaa ggtacgggtg cactgcccaa
     2761 cggcaccgcc ataactgccg cccccgcaac agacgacaaa ccgagttctc cagtcagtga
     2821 caaacttcac gtcagggtcc ccagatggtg cccagcccat ctcacccgaa taagagcttt
     2881 cccgcattag cgaagggctc aagaccttgg gttcttgccg cccaccatgc cccccacctt
     2941 gtttcaacga cctcacagcc cgcctcacaa gcgtcttcca ttcaagactc gggaacagcc
     3001 gccattttgc tgcgctcccc ccaaccccca gttcagggca accttgctcg cggacccaga
     3061 ctacagccct tggcggtctc tccacacgct tccgtcccac cgagcggccc ggcggccacg
     3121 aaagccccgg ccagcccagc agcccgctac tcaccaagtg acgatcacag cgatccacaa
     3181 acaagaaccg cgacccaaat cccggctgcg acggaactag ctgtgccaca cccggcgcgt
     3241 ccttatataa tcatcggcgt tcaccgcccc acggagatcc ctccgcagaa tcgccgagaa
     3301 gggactactt ttcctcgcct gttccgctct ctggaaagaa aaccagtgcc ctagagtcac
     3361 ccaagtcccg tcctaaaatg tccttctgct gatactgggg ttctaaggcc gagtcttatg
     3421 agcagcgggc cgctgtcctg agcgtccggg cggaaggatc aggacgctcc tgcgcccttc
     3481 gtctgacgtg gcagcgctcg ccgtgacgag gggggcgccc gcgggaggcg ccaaaacccg
     3541 gcgcggaggc caggcgcgcc ccgatttcga tattcctcgg atctcggata gatcgggatt
     3601 gaggatgtag gttgaacttc tctcgggtga tcccgatctc ggatattcgg gattggagta
     3661 atacctggcg ataggcgctg ccaagcaata gccaaggcgc aaggccggcc aagagacttg
     3721 cgatagttcg ttcgagaggt agcctgcaag atggatcgga cgaaaggttt gaagattttc
     3781 cgacgagggc cttggagagt tcggaaattt aaatgcaact atggatgaac gaaatagaca
     3841 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3901 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3961 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4021 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4081 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4141 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4201 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4261 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4321 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4381 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4441 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4501 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4561 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4621 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4681 ctggcctttt gctcacatga tttaaatgaa actcgtaatc ggagaacggg tgcaatagca
     4741 aggaccagat cgaacttgac tgacgcaatt ctgagcgtag tcgaaaccca gacaagtcga
     4801 tctcgccaag tagctgactt gactggttaa ttaagtttcg atagcccaag gtaaccaatg
     4861 agattaggga gggaagaaag tttgatgaac tacttctgac ccctagtggt gtccagaaaa
     4921 gaccattaaa ggaatgtctt gatcgaaatc gagagatcgc ctgcaggata gctgacgacc
     4981 ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg gggaagccgt
     5041 atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc atcaggcatt
     5101 tttttctgcg ccaatgccgg tagatagcaa tccggcccga ggggcacaaa aaacccctca
     5161 agacccgttt agaggcccca aggggttatg ctacgatacc gatagagatg ggccaatacc
     5221 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     5281 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5341 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5401 ggtttgtcca aactcatcaa tgtatcttat catgtctgtg agctgaaggt acgctgtatc
     5461 tcagtcagtc aagctagctt acgcataatc tggtacgtca taggggtacg actgaaagta
     5521 aaggttttct tgcagggtgg ctcggttgtc taacatctct tcgcaaagtt tgttgatgtc
     5581 tactgctcct tgcatgctca gtagaacaga tagcaagctg accatcttct cgaaagcttc
     5641 cgtagtatct ttcgcgagca atatatcatt gtgcagctgc acgcactggg cccagagctt
     5701 tgacgaactc tccacccgca gctgttgaag cactgataac agaactacgg atgtacactt
     5761 gacgtcagac attttgctca tggtggcggc cgctggcggg tgtgtcgagt ggatggtagg
     5821 atcgacaaag atctgatctg acggttcact aaaccagctc tgcttatata gacctcccac
     5881 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     5941 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     6001 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcagc
     6061 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     6121 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     6181 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     6241 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     6301 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     6361 acgcggaact ccatatatgg gctatgaact aatga
//
