LOCUS       sequence-295355-        8498 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 8498)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8498)
  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-m4134467
FEATURES             Location/Qualifiers
     source          1..8498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1922..1942
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1943..1969
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2038..2159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2049..2068)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2103..2122
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2274..2320
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2350..2486
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2531..2602
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2828..3416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2908..2927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3916..5125
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4962..4981
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5129..6157
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6184..6232
                     /note="synthetic polyadenylation signal"
     CDS             join(6498..6566,6567..7358)
                     /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(6716..6735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7371..7507
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7652..7723
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7728..7774)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7792..7840
                     /note="synthetic polyadenylation signal"
     enhancer        7921..8224
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8226..8428
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8379..8399
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8497..8498,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggcc tacacaagat atgtggataa caatttttgt gggccggacg gatacccttt
       61 agaatgcatc aaagatttgc ttgcaagggc gggcaaggct tcgtgcactc tgtccgagca
      121 gctggacttc attgatacca agcgcggtgt ttattgttgc cgagagcatg aacacgagat
      181 agcctggtat acggaacggt cagagaaaag ttacgaactc caaaccccat tcgagatcaa
      241 gctagctaaa aagtttgaca ctttcaacgg cgaatgtccc aattttgtct ttcccctgaa
      301 ctctattatc aaaacaatac agcctcgggt ggagaagaag aaattagacg gattcatggg
      361 gcgtattagg agcgtatacc cagtggccag cccgaatgaa tgcaaccaga tgtgtctttc
      421 cacactgatg aagtgcgatc actgtggcga gacttcttgg caaaccggtg acttcgtaaa
      481 agcgacatgc gagttttgtg gtaccgagaa tctgactaag gagggagcta ccacatgcgg
      541 ctatttgcca cagaatgcag tggtcaagat ctactgtcct gcatgccata actctgaagt
      601 cggccccgag cactcgctcg ccgaatatca caatgagagc gggctaaaaa ctatactgag
      661 gaaaggaggt cgcaccatcg cctttggggg ctgcgttttc tcctacgtgg ggtgtcataa
      721 caaatgtgcc tattgggttc ccagagcctc cgctaacatt gggtgcaatc atacgggagt
      781 ggtcggggaa ggctcagaag gcttgaacga taacctcctt gaaattctgc aaaaggagaa
      841 ggtgaatatc aatatcgttg gcgacttcaa gttaaatgag gaaatcgcga ttattctggc
      901 aagtttttct gcctcaacta gcgcctttgt agagacagtg aaaggactcg attataaagc
      961 tttcaaacag atagtcgaga gttgtggtaa cttcaaggtt acaaagggca aggctaaaaa
     1021 gggtgcttgg aacattggag agcagaagtc aatcttgagt ccactgtacg cgtttgcatc
     1081 tgaggccgcc cgtgtggtca ggagcatatt cagtcgaacc ctagaaaccg cacagaatag
     1141 cgtgcgggta ttacagaagg cggctattac tattctcgac ggtatctcgc agtattctct
     1201 gagactgatc gacgccatga tgtttacaag cgatctggca acgaataact tagtggtcat
     1261 ggcgtacata accggagggg tcgtgcaact tacctcccag tggctaacga acatctttgg
     1321 gacggtttac gagaagctaa aacctgtgct ggactggctc gaagagaaat tcaaagaagg
     1381 cgtggaattt ttgcgtgacg ggtgggagat cgtaaagttc atatcaacat gtgcttgcga
     1441 aatcgttggc ggacaaatcg taacttgcgc aaaagagatt aaagaatcgg tgcagacctt
     1501 ctttaaactt gttaacaaat tcctggcttt gtgtgccgat tctattatca tcgggggagc
     1561 aaagcttaaa gccctaaatt taggcgagac atttgtgacg cacagtaagg gcctgtatag
     1621 aaagtgcgtg aagagtcgcg aagaaaccgg actgctcatg cccttaaaag cgccaaaaga
     1681 aataattttc ctggaagggg agacattgcc cactgaagtg ctcactgagg aagtagttct
     1741 gaaaactggg gatctgcaac ctttagagca gccgacctcc gaggccgtcg aggcacctct
     1801 cgtgggaacc ccggtctgta ttaatgggtt gatgctgctc gagattaagg ataccgaaaa
     1861 atactgcgca ctagccccca acatgatggt gactaacaac acatttaccc tcaagggagg
     1921 ggaaaacctt tactttcagt cgtaccccta tgacgtacca gattatgcgt aaggatccgc
     1981 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
     2041 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2101 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2161 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     2221 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
     2281 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
     2341 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     2401 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     2461 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
     2521 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
     2581 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
     2641 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
     2701 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     2761 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2821 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2881 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2941 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3001 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3061 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3121 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3181 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3241 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3301 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3361 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3421 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3481 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3541 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3601 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     3661 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     3721 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     3781 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     3841 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     3901 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     3961 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     4021 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     4081 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     4141 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     4201 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     4261 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     4321 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     4381 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     4441 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     4501 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     4561 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     4621 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     4681 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     4741 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     4801 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     4861 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     4921 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     4981 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     5041 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     5101 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     5161 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     5221 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     5281 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     5341 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     5401 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     5461 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     5521 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     5581 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     5641 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     5701 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     5761 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     5821 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     5881 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     5941 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     6001 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     6061 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     6121 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     6181 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     6241 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     6301 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     6361 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     6421 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     6481 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6541 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6601 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6661 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     6721 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6781 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     6841 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6901 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6961 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7021 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     7081 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     7141 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7201 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7261 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7321 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     7381 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     7441 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     7501 tccttaagat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     7561 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     7621 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     7681 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     7741 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     7801 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     7861 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     7921 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     7981 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     8041 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     8101 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     8161 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     8221 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     8281 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     8341 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     8401 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
     8461 gatcctacca tccactcgac acacccgcca gcggccgc
//
