LOCUS       Exported                8510 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133625
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8510)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8510)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8510
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1916..1936
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1937..1981
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    2050..2171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2061..2080)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2115..2134
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2286..2332
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2362..2498
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2543..2614
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2840..3428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2920..2939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3928..5137
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4974..4993
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5141..6169
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6196..6244
                     /note="synthetic polyadenylation signal"
     CDS             6510..7370
                     /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(6728..6747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7383..7519
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7664..7735
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      7740..7786
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7804..7852
                     /note="synthetic polyadenylation signal"
     enhancer        7933..8236
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8238..8440
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8391..8411
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac actcgctatg tcgataacaa cttctgtggc cctgatggct accctcttga
       61 gtgcattaaa gaccttctag cacgtgctgg taaagcttca tgcactttgt ccgaacaact
      121 ggactttatt gacactaaga ggggtgtata ctgctgccgt gaacatgagc atgaaattgc
      181 ttggtacacg gaacgttctg aaaagagcta tgaattgcag acaccttttg aaattaaatt
      241 ggcaaagaaa tttgacacct tcaatgggga atgtccaaat tttgtatttc ccttaaattc
      301 cataatcaag actattcaac caagggttga aaagaaaaag cttgatggct ttatgggtag
      361 aattcgatct gtctatccag ttgcgtcacc aaatgaatgc aaccaaatgt gcctttcaac
      421 tctcatgaag tgtgatcatt gtggtgaaac ttcatggcag acgggcgatt ttgttaaagc
      481 cacttgcgaa ttttgtggca ctgagaattt gactaaagaa ggtgccacta cttgtggtta
      541 cttaccccaa aatgctgttg ttaaaattta ttgtccagca tgtcacaatt cagaagtagg
      601 acctgagcat agtcttgccg aataccataa tgaatctggc ttgaaaacca ttcttcgtaa
      661 gggtggtcgc actattgcct ttggaggctg tgtgttctct tatgttggtt gccataacaa
      721 gtgtgcctat tgggttccac gtgctagcgc taacataggt tgtaaccata caggtgttgt
      781 tggagaaggt tccgaaggtc ttaatgacaa ccttcttgaa atactccaaa aagagaaagt
      841 caacatcaat attgttggtg actttaaact taatgaagag atcgccatta ttttggcatc
      901 tttttctgct tccacaagtg cttttgtgga aactgtgaaa ggtttggatt ataaagcatt
      961 caaacaaatt gttgaatcct gtggtaattt taaagttaca aaaggaaaag ctaaaaaagg
     1021 tgcctggaat attggtgaac agaaatcaat actgagtcct ctttatgcat ttgcatcaga
     1081 ggctgctcgt gttgtacgat caattttctc ccgcactctt gaaactgctc aaaattctgt
     1141 gcgtgtttta cagaaggccg ctataacaat actagatgga atttcacagt attcactgag
     1201 actcattgat gctatgatgt tcacatctga tttggctact aacaatctag ttgtaatggc
     1261 ctacattaca ggtggtgttg ttcagttgac ttcgcagtgg ctaactaaca tctttggcac
     1321 tgtttatgaa aaactcaaac ccgtccttga ttggcttgaa gagaagttta aggaaggtgt
     1381 agagtttctt agagacggtt gggaaattgt taaatttatc tcaacctgtg cttgtgaaat
     1441 tgtcggtgga caaattgtca cctgtgcaaa ggaaattaag gagagtgttc agacattctt
     1501 taagcttgta aataaatttt tggctttgtg tgctgactct atcattattg gtggagctaa
     1561 acttaaagcc ttgaatttag gtgaaacatt tgtcacgcac tcaaagggat tgtacagaaa
     1621 gtgtgttaaa tccagagaag aaactggcct actcatgcct ctaaaagccc caaaagaaat
     1681 tatcttctta gagggagaaa cacttcccac agaagtgtta acagaggaag ttgtcttgaa
     1741 aactggtgat ttacaaccat tagaacaacc tactagtgaa gctgttgaag ctccattggt
     1801 tggtacacca gtttgtatta acgggcttat gttgctcgaa atcaaagaca cagaaaagta
     1861 ctgtgccctt gcacctaata tgatggtaac aaacaatacc ttcacactca aaggcgaaaa
     1921 cctttacttt cagtcgaaag aaaccgcggc agctaagttc gagaggcagc acatggacag
     1981 ttaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     2041 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2101 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2161 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2221 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     2281 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     2341 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     2401 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     2461 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     2521 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     2581 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     2641 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     2701 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     2761 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2821 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2881 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2941 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3001 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3061 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3121 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3181 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3241 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3301 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3361 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3421 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3541 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3661 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     3721 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     3781 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     3841 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     3901 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     3961 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     4021 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     4081 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     4141 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     4201 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     4261 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     4321 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     4381 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     4441 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     4501 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     4561 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     4621 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     4681 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     4741 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     4801 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     4861 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     4921 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     4981 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     5041 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     5101 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     5161 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     5221 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     5281 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     5341 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     5401 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     5461 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     5521 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     5581 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     5641 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     5701 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     5761 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     5821 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     5881 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     5941 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     6001 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     6061 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     6121 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     6181 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     6241 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     6301 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     6361 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     6421 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     6481 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     6541 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6601 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     6661 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     6721 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6781 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     6841 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6901 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     6961 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7021 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     7081 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     7141 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7201 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7261 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7321 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     7381 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     7441 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     7501 accagaaatt tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     7561 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     7621 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     7681 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     7741 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     7801 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     7861 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     7921 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7981 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     8041 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     8101 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     8161 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     8221 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     8281 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     8341 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     8401 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     8461 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
