LOCUS       sequence-338673-        8075 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 8075)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8075)
  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-m4134534
FEATURES             Location/Qualifiers
     source          1..8075
                     /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             2521..2562
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2615..2736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2626..2645)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2680..2699
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2851..2897
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2927..3063
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3108..3179
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3405..3993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3485..3504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4493..5702
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5539..5558
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5706..6305
                     /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(5706..5725)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6202..6222
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6332..6380
                     /note="synthetic polyadenylation signal"
     CDS             join(6646..6714,6715..7506)
                     /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(6864..6883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7519..7655
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7807..7878
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7883..7929)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7947..7995
                     /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 gggaaaccta tacccaatcc acttctcggc ttagattcta cctaagctag cttgactgac
     2581 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
     2641 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2701 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2761 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2821 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
     2881 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
     2941 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     3001 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     3061 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
     3121 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
     3181 tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg
     3241 agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt
     3301 ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc
     3361 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3421 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3481 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3541 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3601 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3661 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3721 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3781 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3841 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3901 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3961 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4021 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4081 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4141 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4201 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
     4261 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
     4321 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
     4381 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
     4441 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg
     4501 cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca
     4561 gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
     4621 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
     4681 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
     4741 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
     4801 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
     4861 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
     4921 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
     4981 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
     5041 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
     5101 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc
     5161 gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt
     5221 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     5281 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     5341 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
     5401 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     5461 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     5521 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     5581 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     5641 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     5701 acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     5761 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5821 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5881 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5941 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     6001 gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg caccggccca
     6061 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     6121 tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg gtgcccgcct
     6181 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     6241 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     6301 cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat
     6361 gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc
     6421 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     6481 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     6541 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     6601 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     6661 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6721 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     6781 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     6841 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6901 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     6961 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7021 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     7081 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7141 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7201 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     7261 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7321 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7381 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7441 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7501 tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     7561 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     7621 catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact
     7681 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     7741 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     7801 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     7861 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     7921 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
     7981 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
     8041 gggtcattag ttcatagccc atatatggag ttccg
//
