LOCUS       sequence-295599-        8492 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8492)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8492)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133276
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1916..1936
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1937..1963
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2032..2153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2043..2062)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2097..2116
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2268..2314
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2344..2480
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2525..2596
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2822..3410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2902..2921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3910..5119
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4956..4975
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5123..6151
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6178..6226
                     /note="synthetic polyadenylation signal"
     CDS             join(6492..6560,6561..7352)
                     /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(6710..6729)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7365..7501
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7646..7717
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7722..7768)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7786..7834
                     /note="synthetic polyadenylation signal"
     enhancer        7915..8218
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8220..8422
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8373..8393
                     /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 cagtcgtacc cctatgacgt accagattat gcgtaaggat ccgcaggcct
     1981 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     2041 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2101 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2161 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2221 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     2281 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     2341 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     2401 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     2461 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     2521 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     2581 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     2641 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     2701 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     2761 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3001 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3241 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     3661 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     3721 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     3781 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     3841 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     3901 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     3961 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     4021 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     4081 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     4141 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     4201 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     4261 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     4321 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     4381 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     4441 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     4501 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     4561 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     4621 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     4681 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     4741 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt
     4801 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     4861 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     4921 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     4981 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     5041 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     5101 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     5161 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     5221 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     5281 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     5341 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     5401 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     5461 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     5521 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     5581 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     5641 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     5701 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     5761 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     5821 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     5881 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     5941 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     6001 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     6061 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     6121 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     6181 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     6241 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     6301 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     6361 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     6421 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     6481 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6541 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6601 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6661 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6721 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6781 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6841 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6901 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6961 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7021 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     7081 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     7141 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7201 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7261 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7321 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     7381 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     7441 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     7501 agatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     7561 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     7621 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     7681 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     7741 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     7801 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     7861 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     7921 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     7981 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     8041 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     8101 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     8161 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
     8221 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     8281 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     8341 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     8401 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
     8461 accatccact cgacacaccc gccagcggcc gc
//
