LOCUS       sequence-295722-        8483 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 8483)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8483)
  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-m4133423
FEATURES             Location/Qualifiers
     source          1..8483
                     /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..1954
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2023..2144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2034..2053)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2088..2107
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2259..2305
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2335..2471
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2516..2587
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2813..3401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2893..2912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3901..5110
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4947..4966
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5114..6142
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6169..6217
                     /note="synthetic polyadenylation signal"
     CDS             join(6483..6551,6552..7343)
                     /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(6701..6720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7356..7492
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7637..7708
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7713..7759)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7777..7825
                     /note="synthetic polyadenylation signal"
     enhancer        7906..8209
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8211..8413
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8364..8384
                     /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 cagtcgcacc atcaccatca ccattaagga tccgcaggcc tctgctagct
     1981 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
     2041 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2101 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2161 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     2221 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
     2281 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
     2341 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     2401 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     2461 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
     2521 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
     2581 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
     2641 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     2701 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     2761 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3001 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3241 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3601 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     3661 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     3721 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     3781 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     3841 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     3901 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg
     3961 ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc
     4021 ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg
     4081 acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga
     4141 aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg
     4201 attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc
     4261 gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg
     4321 gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc
     4381 aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc
     4441 agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag
     4501 gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc
     4561 gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga
     4621 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     4681 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     4741 gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg
     4801 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     4861 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     4921 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     4981 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
     5041 atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt
     5101 tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     5161 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     5221 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     5281 tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc tcccgattcc
     5341 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     5401 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     5461 cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg ggttcggccc
     5521 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     5581 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     5641 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     5701 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     5761 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     5821 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     5881 gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg accaactcta
     5941 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     6001 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgccgc
     6061 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     6121 tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa taaaatcgct
     6181 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg
     6241 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     6301 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     6361 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     6421 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     6481 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6541 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6601 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6661 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6721 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6781 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6841 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6901 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6961 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7021 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     7081 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     7141 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7201 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7261 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7321 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     7381 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     7441 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt
     7501 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     7561 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     7621 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     7681 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     7741 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     7801 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     7861 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     7921 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     7981 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     8041 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     8101 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     8161 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     8221 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     8281 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     8341 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     8401 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     8461 tcgacacacc cgccagcggc cgc
//
