LOCUS       Exported                6758 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252855
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6758)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6758)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6758
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    85..156
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      161..207
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    225..273
                     /note="synthetic polyadenylation signal"
     enhancer        354..657
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        659..861
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     812..832
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1188..1208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1557..1574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1651..1772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1662..1681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1716..1735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1887..1933
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1963..2099
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2144..2215
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2441..3029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2521..2540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3529..4738
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4575..4594
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4742..5341
                     /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(4742..4761)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5238..5258
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5368..5416
                     /note="synthetic polyadenylation signal"
     CDS             5682..6542
                     /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(5900..5919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6555..6691
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt
       61 tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc
      121 ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc
      181 gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac
      241 acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag
      301 atctagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      361 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      421 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      481 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      541 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      601 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg
      661 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      721 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      781 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      841 gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta
      901 ccatccactc gacacacccg ccagcggccg ccaccatgag caaaatgtct gacgtcaagt
      961 gtacatccgt agttctgtta tcagtgcttc aacagctgcg ggtggagagt tcgtcaaagc
     1021 tctgggccca gtgcgtgcag ctgcacaatg atatattgct cgcgaaagat actacggaag
     1081 ctttcgagaa gatggtcagc ttgctatctg ttctactgag catgcaagga gcagtagaca
     1141 tcaacaaact ttgcgaagag atgttagaca accgagccac cctgcaagaa aacctttact
     1201 ttcagtcgat gagtgacaaa atcattcatc taaccgatga ctcctttgat acagacgtgt
     1261 tgaaggccga tggtgcgata ctggtagatt tctgggctga atggtgtggc ccgtgcaaga
     1321 tgatcgcacc cattctggac gagatcgccg acgagtatca aggaaaatta actgtcgcta
     1381 agcttaatat agatcagaac ccagggacgg cacctaaata cggcatcagg ggaataccta
     1441 ccctcctgct ctttaagaat ggggaagttg ccgcgacaaa agtgggagcc ctgtctaagg
     1501 gccagctaaa agagttcttg gacgctaacc tggcaggttc aggctcgggg cacatgcacc
     1561 atcaccatca ccatagcagc gggctcgtgc cacgcggcta agctagcttg actgactgag
     1621 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
     1681 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1741 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1801 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     1861 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
     1921 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
     1981 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     2041 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     2101 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
     2161 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
     2221 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
     2281 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
     2341 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
     2401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2881 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2941 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3001 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3061 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3121 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3181 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3241 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
     3301 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
     3361 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
     3421 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
     3481 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc
     3541 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg
     3601 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     3661 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     3721 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     3781 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     3841 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     3901 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     3961 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     4021 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     4081 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     4141 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga
     4201 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt
     4261 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     4321 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     4381 tctgttggct tataatgcag ggtggggcca ccagccggta ggtgtgcggt aggcttttct
     4441 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     4501 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     4561 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     4621 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     4681 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa
     4741 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     4801 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     4861 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4921 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4981 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5041 ttcccggctg gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga
     5101 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5161 cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct
     5221 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     5281 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta
     5341 ggctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt
     5401 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag
     5461 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     5521 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     5581 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     5641 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     5701 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5761 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     5821 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     5881 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5941 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     6001 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6061 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     6121 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6181 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6241 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     6301 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6361 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6421 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6481 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6541 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     6601 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     6661 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     6721 ctctatcgaa tctccgtcgt ttcgagctta cgcgaaca
//
