LOCUS       sequence-320774-        4592 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 4592)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4592)
  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-m4134392
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(9..1218)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(153..172)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2532..2603
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2648..2784)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2797..3588,3589..3657))
                     /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     3420..3439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    3923..3971
                     /note="synthetic polyadenylation signal"
     CDS             complement(join(3998..4592,1..5))
                     /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(4081..4101)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     join(4578..4592,1..5)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gtcatgttgt ctaacaaaaa agccaaaaac ggccagaatt tagcggacaa tttacaagtc
       61 taacactgaa aattacatat tgacccaaat gattacattt caaaaggtgc ctaaaaaact
      121 tcacaaaaca cactcgccaa ccccgagcgc atagttcaaa accggagctt cagctactta
      181 agaagatagg tacataaaac cgaccaaaga aactgacgcc tcacttatcc ctcccctcac
      241 cagaggtccg gcgcctgtcg attcaggaga gcctacccta ggcccgaacc ctgcgtcctg
      301 cgacggagaa aagcctaccg cacacctacc ggcaggtggc cccaccctgc attataagcc
      361 aacagaacgg gtgacgtcac gacacgacga gggcgcgcgc tcccaaaggt acgggtgcac
      421 tgcccaacgg caccgccata actgccgccc ccgcaacaga cgacaaaccg agttctccag
      481 tcagtgacaa acttcacgtc agggtcccca gatggtgccc agcccatctc acccgaataa
      541 gagctttccc gcattagcga agggctcaag accttgggtt cttgccgccc accatgcccc
      601 ccaccttgtt tcaacgacct cacagcccgc ctcacaagcg tcttccattc aagactcggg
      661 aacagccgcc attttgctgc gctcccccca acccccagtt cagggcaacc ttgctcgcgg
      721 acccagacta cagcccttgg cggtctctcc acacgcttcc gtcccaccga gcggcccggc
      781 ggccacgaaa gccccggcca gcccagcagc ccgctactca ccaagtgacg atcacagcga
      841 tccacaaaca agaaccgcga cccaaatccc ggctgcgacg gaactagctg tgccacaccc
      901 ggcgcgtcct tatataatca tcggcgttca ccgccccacg gagatccctc cgcagaatcg
      961 ccgagaaggg actacttttc ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta
     1021 gagtcaccca agtcccgtcc taaaatgtcc ttctgctgat actggggttc taaggccgag
     1081 tcttatgagc agcgggccgc tgtcctgagc gtccgggcgg aaggatcagg acgctcctgc
     1141 gcccttcgtc tgacgtggca gcgctcgccg tgacgagggg ggcgcccgcg ggaggcgcca
     1201 aaacccggcg cggaggccag gcgcgccccg atttcgatat tcctcggatc tcggatagat
     1261 cgggattgag gatgtaggtt gaacttctct cgggtgatcc cgatctcgga tattcgggat
     1321 tggagtaata cctggcgata ggcgctgcca agcaatagcc aaggcgcaag gccggccaag
     1381 agacttgcga tagttcgttc gagaggtagc ctgcaagatg gatcggacga aaggtttgaa
     1441 gattttccga cgagggcctt ggagagttcg gaaatttaaa tgcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgattt aaatgaaact cgtaatcgga gaacgggtgc
     2401 aatagcaagg accagatcga acttgactga cgcaattctg agcgtagtcg aaacccagac
     2461 aagtcgatct cgccaagtag ctgacttgac tggttaatta agtttcgata gcccaaggta
     2521 accaatgaga ttagggaggg aagaaagttt gatgaactac ttctgacccc tagtggtgtc
     2581 cagaaaagac cattaaagga atgtcttgat cgaaatcgag agatcgcctg caggatagct
     2641 gacgacctta aggataaatt tctggtaagg aggacacgta tggaagtggg caagttgggg
     2701 aagccgtatc cgttgctgaa tctggcatat gtgggagtat aagacgcgca gcgtcgcatc
     2761 aggcattttt ttctgcgcca atgcacctag aatcggttac caatgcttaa tcagtgaggc
     2821 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2881 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2941 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3001 cagaagtggt cctgcaactt tatccgcctc catccagtct atcaactgtt gccgggaagc
     3061 tagagtaagt agttcgccag ttaatagttt acgcaaggtt gttgccattg ctacaggcat
     3121 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3181 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccaat
     3241 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3301 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgaat actcaaccaa
     3361 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3421 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaagc gttcttcggg
     3481 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgcgc
     3541 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3601 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatgcg
     3661 aaagcctcct gctgatcctg tctcttgata gatgtttgca aaagctagga ctcgattata
     3721 cgttgaccga tcagactatc aatatctagg tttaaacaga tcggtacgct gccgtgacca
     3781 ggcttgagta caatatctga cgattggact tcctgggtcc caagctggat tccgactggc
     3841 tgactgccaa gcaacgatat ggatcgtcga gactcgcact gagaggatcg ttctcggtac
     3901 accaggcgcg cctagcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     3961 gcgattttat tatctaagac taggcttaca tactagccta ggcaccgggc ttgcgggtca
     4021 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
     4081 gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc
     4141 gctcggcggc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
     4201 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     4261 ggaggccctc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     4321 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     4381 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     4441 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
     4501 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
     4561 gggtggcgag gcgcaccgtg ggcttgtact cg
//
