LOCUS       sequence-311988-        8423 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 8423)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8423)
  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-m4134436
FEATURES             Location/Qualifiers
     source          1..8423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2500..2520
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2869..2886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2963..3084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3028..3047
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3199..3245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3275..3411
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3456..3527
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3753..4341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3833..3852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4841..6050
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5887..5906
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6054..6653
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6054..6073)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6550..6570
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6680..6728
                     /note="synthetic polyadenylation signal"
     CDS             join(6994..7062,7063..7854)
                     /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(7212..7231)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7867..8003
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8155..8226
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8231..8277)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8295..8343
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggccta cacaagatat
      601 gtggataaca atttttgtgg gccggacgga taccctttag aatgcatcaa agatttgctt
      661 gcaagggcgg gcaaggcttc gtgcactctg tccgagcagc tggacttcat tgataccaag
      721 cgcggtgttt attgttgccg agagcatgaa cacgagatag cctggtatac ggaacggtca
      781 gagaaaagtt acgaactcca aaccccattc gagatcaagc tagctaaaaa gtttgacact
      841 ttcaacggcg aatgtcccaa ttttgtcttt cccctgaact ctattatcaa aacaatacag
      901 cctcgggtgg agaagaagaa attagacgga ttcatggggc gtattaggag cgtataccca
      961 gtggccagcc cgaatgaatg caaccagatg tgtctttcca cactgatgaa gtgcgatcac
     1021 tgtggcgaga cttcttggca aaccggtgac ttcgtaaaag cgacatgcga gttttgtggt
     1081 accgagaatc tgactaagga gggagctacc acatgcggct atttgccaca gaatgcagtg
     1141 gtcaagatct actgtcctgc atgccataac tctgaagtcg gccccgagca ctcgctcgcc
     1201 gaatatcaca atgagagcgg gctaaaaact atactgagga aaggaggtcg caccatcgcc
     1261 tttgggggct gcgttttctc ctacgtgggg tgtcataaca aatgtgccta ttgggttccc
     1321 agagcctccg ctaacattgg gtgcaatcat acgggagtgg tcggggaagg ctcagaaggc
     1381 ttgaacgata acctccttga aattctgcaa aaggagaagg tgaatatcaa tatcgttggc
     1441 gacttcaagt taaatgagga aatcgcgatt attctggcaa gtttttctgc ctcaactagc
     1501 gcctttgtag agacagtgaa aggactcgat tataaagctt tcaaacagat agtcgagagt
     1561 tgtggtaact tcaaggttac aaagggcaag gctaaaaagg gtgcttggaa cattggagag
     1621 cagaagtcaa tcttgagtcc actgtacgcg tttgcatctg aggccgcccg tgtggtcagg
     1681 agcatattca gtcgaaccct agaaaccgca cagaatagcg tgcgggtatt acagaaggcg
     1741 gctattacta ttctcgacgg tatctcgcag tattctctga gactgatcga cgccatgatg
     1801 tttacaagcg atctggcaac gaataactta gtggtcatgg cgtacataac cggaggggtc
     1861 gtgcaactta cctcccagtg gctaacgaac atctttggga cggtttacga gaagctaaaa
     1921 cctgtgctgg actggctcga agagaaattc aaagaaggcg tggaattttt gcgtgacggg
     1981 tgggagatcg taaagttcat atcaacatgt gcttgcgaaa tcgttggcgg acaaatcgta
     2041 acttgcgcaa aagagattaa agaatcggtg cagaccttct ttaaacttgt taacaaattc
     2101 ctggctttgt gtgccgattc tattatcatc gggggagcaa agcttaaagc cctaaattta
     2161 ggcgagacat ttgtgacgca cagtaagggc ctgtatagaa agtgcgtgaa gagtcgcgaa
     2221 gaaaccggac tgctcatgcc cttaaaagcg ccaaaagaaa taattttcct ggaaggggag
     2281 acattgccca ctgaagtgct cactgaggaa gtagttctga aaactgggga tctgcaacct
     2341 ttagagcagc cgacctccga ggccgtcgag gcacctctcg tgggaacccc ggtctgtatt
     2401 aatgggttga tgctgctcga gattaaggat accgaaaaat actgcgcact agcccccaac
     2461 atgatggtga ctaacaacac atttaccctc aagggagggg aaaaccttta ctttcagtcg
     2521 atgagtgaca aaatcattca tctaaccgat gactcctttg atacagacgt gttgaaggcc
     2581 gatggtgcga tactggtaga tttctgggct gaatggtgtg gcccgtgcaa gatgatcgca
     2641 cccattctgg acgagatcgc cgacgagtat caaggaaaat taactgtcgc taagcttaat
     2701 atagatcaga acccagggac ggcacctaaa tacggcatca ggggaatacc taccctcctg
     2761 ctctttaaga atggggaagt tgccgcgaca aaagtgggag ccctgtctaa gggccagcta
     2821 aaagagttct tggacgctaa cctggcaggt tcaggctcgg ggcacatgca ccatcaccat
     2881 caccatagca gcgggctcgt gccacgcggc taagctagct tgactgactg agatacagcg
     2941 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3001 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3061 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3121 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     3181 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     3241 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     3301 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     3361 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     3421 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     3481 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     3541 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     3601 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     3661 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     3721 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3781 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3841 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3901 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3961 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4021 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4081 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4141 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4201 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4261 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4321 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4381 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4441 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4501 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4561 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     4621 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     4681 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     4741 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     4801 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg
     4861 gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc
     4921 aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
     4981 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
     5041 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
     5101 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
     5161 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
     5221 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
     5281 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
     5341 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
     5401 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     5461 ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt
     5521 attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     5581 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt
     5641 gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg
     5701 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     5761 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     5821 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     5881 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     5941 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     6001 cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgaccg
     6061 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     6121 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     6181 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     6241 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     6301 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     6361 tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag gagcccgcgt
     6421 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     6481 tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc ctggagacct
     6541 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     6601 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc taggctagta
     6661 tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt
     6721 tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc
     6781 tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga
     6841 agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa
     6901 cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc
     6961 tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc
     7021 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7081 aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc
     7141 aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact
     7201 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     7261 ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag
     7321 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7381 aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt
     7441 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7501 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt
     7561 aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg
     7621 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7681 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7741 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7801 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt
     7861 ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     7921 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     7981 ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg
     8041 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     8101 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     8161 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     8221 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     8281 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     8341 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
     8401 catagcccat atatggagtt ccg
//
