LOCUS       Exported                7220 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133802
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7220)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7220)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7220
                     /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             1618..1638
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1642..1707
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1760..1881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1771..1790)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1825..1844
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1996..2042
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2072..2208
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2253..2324
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2550..3138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2630..2649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3638..4847
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4684..4703
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4851..5450
                     /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(4851..4870)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5347..5367
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5477..5525
                     /note="synthetic polyadenylation signal"
     CDS             5791..6651
                     /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(6009..6028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6664..6800
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6952..7023
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      7028..7074
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7092..7140
                     /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 ccatgttaga aaatgtggct
      601 tttaatgttg taaataaggg acactttgat ggacaacagg gtgaagtacc agtttctatc
      661 attaataaca ctgtttacac aaaagttgat ggtgttgatg tagaattgtt tgaaaataaa
      721 acaacattac ctgttaatgt agcatttgag ctttgggcta agcgcaacat taaaccagta
      781 ccagaggtga aaatactcaa taatttgggt gtggacattg ctgctaatac tgtgatctgg
      841 gactacaaaa gagatgctcc agcacatata tctactattg gtgtttgttc tatgactgac
      901 atagccaaga aaccaactga aacgatttgt gcaccactca ctgtcttttt tgatggtaga
      961 gttgatggtc aagtagactt atttagaaat gcccgtaatg gtgttcttat tacagaaggt
     1021 agtgttaaag gtttacaacc atctgtaggt cccaaacaag ctagtcttaa tggagtcaca
     1081 ttaattggag aagccgtaaa aacacagttc aattattata agaaagttga tggtgttgtc
     1141 caacaattac ctgaaactta ctttactcag agtagaaatt tacaagaatt taaacccagg
     1201 agtcaaatgg aaattgattt cttagaatta gctatggatg aattcattga acggtataaa
     1261 ttagaaggct atgccttcga acatatcgtt tatggagatt ttagtcatag tcagttaggt
     1321 ggtttacatc tactgattgg actagctaaa cgttttaagg aatcaccttt tgaattagaa
     1381 gattttattc ctatggacag tacagttaaa aactatttca taacagatgc gcaaacaggt
     1441 tcatctaagt gtgtgtgttc tgttattgat ttattacttg atgattttgt tgaaataata
     1501 aaatcccaag atttatctgt agtttctaag gttgtcaaag tgactattga ctatacagaa
     1561 atttcattta tgctttggtg taaagatggc catgtagaaa cattttaccc aaaattagaa
     1621 aacctttact ttcagtcgat ggattataag gaccacgatg gagactacaa agatcatgac
     1681 attgattaca aggacgacga tgacaaataa gctagcttga ctgactgaga tacagcgtac
     1741 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1801 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1861 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1921 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     1981 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2041 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     2101 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     2161 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     2221 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     2281 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     2341 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     2401 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     2461 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     2521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3361 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     3421 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     3481 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     3541 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     3601 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg
     3661 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     3721 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     3781 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     3841 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     3901 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     3961 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     4021 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     4081 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     4141 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     4201 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     4261 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     4321 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     4381 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     4441 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     4501 ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga
     4561 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     4621 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     4681 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     4741 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt
     4801 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac atgaccgagt
     4861 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4921 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4981 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     5041 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     5101 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     5161 ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     5221 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     5281 tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     5341 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     5401 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag gctagtatgt
     5461 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     5521 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     5581 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     5641 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     5701 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     5761 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     5821 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5881 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     5941 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     6001 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6061 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     6121 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6181 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     6241 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6301 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     6361 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     6421 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6481 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6541 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6601 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     6661 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     6721 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     6781 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     6841 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6901 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     6961 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     7021 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     7081 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
     7141 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
     7201 agcccatata tggagttccg
//
