LOCUS       sequence-295482-        8855 bp    DNA     circular SYN 15-DEC-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 8855)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8855)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134172
FEATURES             Location/Qualifiers
     source          1..8855
                     /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             2285..2302
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2395..2516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2406..2425)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2460..2479
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2631..2677
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2707..2843
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2888..2959
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3185..3773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3265..3284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4273..5482
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5319..5338
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5486..6514
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6541..6589
                     /note="synthetic polyadenylation signal"
     CDS             join(6855..6923,6924..7715)
                     /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(7073..7092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7728..7864
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8009..8080
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8085..8131)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8149..8197
                     /note="synthetic polyadenylation signal"
     enhancer        8278..8581
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8583..8785
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8736..8756
                     /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 cagtcgatga gtgacaaaat cattcatcta accgatgact cctttgatac
     1981 agacgtgttg aaggccgatg gtgcgatact ggtagatttc tgggctgaat ggtgtggccc
     2041 gtgcaagatg atcgcaccca ttctggacga gatcgccgac gagtatcaag gaaaattaac
     2101 tgtcgctaag cttaatatag atcagaaccc agggacggca cctaaatacg gcatcagggg
     2161 aatacctacc ctcctgctct ttaagaatgg ggaagttgcc gcgacaaaag tgggagccct
     2221 gtctaagggc cagctaaaag agttcttgga cgctaacctg gcaggttcag gctcggggca
     2281 catgcaccat caccatcacc atagcagcgg gctcgtgcca cgcggctaag gatccgcagg
     2341 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
     2401 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2461 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2521 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2581 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
     2641 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
     2701 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     2761 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     2821 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
     2881 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     2941 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     3001 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     3061 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     3121 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3901 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     4021 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     4081 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     4141 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     4201 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     4261 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     4321 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     4381 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     4441 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     4501 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     4561 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     4621 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     4681 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     4741 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     4801 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     4861 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     4921 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     4981 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     5041 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     5101 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     5161 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     5221 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     5281 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     5341 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     5401 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     5461 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     5521 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     5581 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     5641 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     5701 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     5761 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     5821 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     5881 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     5941 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     6001 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     6061 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     6121 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     6181 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     6241 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     6301 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     6361 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     6421 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     6481 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     6541 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     6601 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     6661 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     6721 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     6781 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     6841 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6901 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6961 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7021 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7081 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7141 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     7201 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7261 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7321 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7381 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     7441 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     7501 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7561 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7621 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7681 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     7741 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     7801 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     7861 ttaagatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     7921 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     7981 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     8041 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     8101 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     8161 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     8221 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     8281 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     8341 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     8401 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     8461 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     8521 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     8581 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     8641 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     8701 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     8761 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     8821 cctaccatcc actcgacaca cccgccagcg gccgc
//
