LOCUS       sequence-295608-        8489 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 8489)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8489)
  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-m4133983
FEATURES             Location/Qualifiers
     source          1..8489
                     /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..1960
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2029..2150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2040..2059)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2094..2113
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2265..2311
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2341..2477
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2522..2593
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2899..2918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3907..5116
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4953..4972
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5120..6148
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6175..6223
                     /note="synthetic polyadenylation signal"
     CDS             join(6489..6557,6558..7349)
                     /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(6707..6726)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7362..7498
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7643..7714
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7719..7765)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7783..7831
                     /note="synthetic polyadenylation signal"
     enhancer        7912..8215
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8217..8419
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8370..8390
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8488..8489,1..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 cgcaccatca ccatcaccat taaggatccg caggcctctg
     1981 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     2041 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2101 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2161 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2221 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     2281 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     2341 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     2401 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     2461 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     2521 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     2581 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     2641 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     2701 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3241 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3541 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     3661 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     3721 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3781 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3841 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3901 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3961 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     4021 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     4081 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     4141 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     4201 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     4261 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     4321 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     4381 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     4441 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     4501 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     4561 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     4621 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     4681 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     4741 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     4801 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4861 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4921 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4981 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     5041 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     5101 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     5161 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     5221 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     5281 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     5341 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     5401 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     5461 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     5521 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     5581 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     5641 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     5701 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     5761 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     5821 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     5881 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     5941 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     6001 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     6061 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     6121 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     6181 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     6241 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     6301 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     6361 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     6421 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     6481 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6601 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6661 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6721 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6781 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6901 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7021 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     7081 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     7141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7321 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     7381 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     7441 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     7501 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     7561 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     7621 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     7681 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     7741 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     7801 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     7861 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7921 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7981 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8041 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     8101 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     8161 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     8221 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     8281 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     8341 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     8401 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     8461 atccactcga cacacccgcc agcggccgc
//
