LOCUS       sequence-295650-        8861 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 8861)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8861)
  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-m4134527
FEATURES             Location/Qualifiers
     source          1..8861
                     /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             2291..2308
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2401..2522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2412..2431)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2466..2485
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2637..2683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2713..2849
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2894..2965
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3191..3779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3271..3290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4279..5488
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5325..5344
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5492..6520
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6547..6595
                     /note="synthetic polyadenylation signal"
     CDS             join(6861..6929,6930..7721)
                     /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(7079..7098)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7734..7870
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8015..8086
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8091..8137)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8155..8203
                     /note="synthetic polyadenylation signal"
     enhancer        8284..8587
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8589..8791
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8742..8762
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8860..8861,1..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 cgatgagtga caaaatcatt catctaaccg atgactcctt
     1981 tgatacagac gtgttgaagg ccgatggtgc gatactggta gatttctggg ctgaatggtg
     2041 tggcccgtgc aagatgatcg cacccattct ggacgagatc gccgacgagt atcaaggaaa
     2101 attaactgtc gctaagctta atatagatca gaacccaggg acggcaccta aatacggcat
     2161 caggggaata cctaccctcc tgctctttaa gaatggggaa gttgccgcga caaaagtggg
     2221 agccctgtct aagggccagc taaaagagtt cttggacgct aacctggcag gttcaggctc
     2281 ggggcacatg caccatcacc atcaccatag cagcgggctc gtgccacgcg gctaaggatc
     2341 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     2401 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2461 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2521 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2581 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     2641 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     2701 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     2761 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     2821 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     2881 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     2941 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     3001 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     3061 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     3121 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3541 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3601 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3661 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3721 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3781 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3841 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3901 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3961 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     4021 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     4081 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     4141 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     4201 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     4261 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     4321 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     4381 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     4441 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     4501 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     4561 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     4621 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     4681 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     4741 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     4801 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     4861 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     4921 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     4981 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     5041 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     5101 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     5161 cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     5221 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     5281 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     5341 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     5401 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     5461 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     5521 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     5581 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     5641 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     5701 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     5761 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     5821 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     5881 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     5941 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     6001 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     6061 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     6121 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     6181 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     6241 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     6301 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     6361 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     6421 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     6481 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     6541 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     6601 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     6661 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     6721 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     6781 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     6841 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     6901 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6961 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     7021 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     7081 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     7141 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     7201 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     7261 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     7321 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7381 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     7441 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     7501 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7561 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7621 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7681 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     7741 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     7801 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     7861 ttatccttaa gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     7921 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     7981 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     8041 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     8101 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     8161 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     8221 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     8281 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     8341 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     8401 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     8461 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     8521 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     8581 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     8641 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     8701 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     8761 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
     8821 gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
