LOCUS       sequence-310593-        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-m4133544
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..1210)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(145..164)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      1710..2298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2199..2218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2524..2595
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2640..2776)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2789..3580,3581..3649))
                     /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     3412..3431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    3915..3963
                     /note="synthetic polyadenylation signal"
     CDS             complement(3990..4589)
                     /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(4073..4093)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4570..4589
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gtctaacaaa aaagccaaaa acggccagaa tttagcggac aatttacaag tctaacactg
       61 aaaattacat attgacccaa atgattacat ttcaaaaggt gcctaaaaaa cttcacaaaa
      121 cacactcgcc aaccccgagc gcatagttca aaaccggagc ttcagctact taagaagata
      181 ggtacataaa accgaccaaa gaaactgacg cctcacttat ccctcccctc accagaggtc
      241 cggcgcctgt cgattcagga gagcctaccc taggcccgaa ccctgcgtcc tgcgacggag
      301 aaaagcctac cgcacaccta ccggcaggtg gccccaccct gcattataag ccaacagaac
      361 gggtgacgtc acgacacgac gagggcgcgc gctcccaaag gtacgggtgc actgcccaac
      421 ggcaccgcca taactgccgc ccccgcaaca gacgacaaac cgagttctcc agtcagtgac
      481 aaacttcacg tcagggtccc cagatggtgc ccagcccatc tcacccgaat aagagctttc
      541 ccgcattagc gaagggctca agaccttggg ttcttgccgc ccaccatgcc ccccaccttg
      601 tttcaacgac ctcacagccc gcctcacaag cgtcttccat tcaagactcg ggaacagccg
      661 ccattttgct gcgctccccc caacccccag ttcagggcaa ccttgctcgc ggacccagac
      721 tacagccctt ggcggtctct ccacacgctt ccgtcccacc gagcggcccg gcggccacga
      781 aagccccggc cagcccagca gcccgctact caccaagtga cgatcacagc gatccacaaa
      841 caagaaccgc gacccaaatc ccggctgcga cggaactagc tgtgccacac ccggcgcgtc
      901 cttatataat catcggcgtt caccgcccca cggagatccc tccgcagaat cgccgagaag
      961 ggactacttt tcctcgcctg ttccgctctc tggaaagaaa accagtgccc tagagtcacc
     1021 caagtcccgt cctaaaatgt ccttctgctg atactggggt tctaaggccg agtcttatga
     1081 gcagcgggcc gctgtcctga gcgtccgggc ggaaggatca ggacgctcct gcgcccttcg
     1141 tctgacgtgg cagcgctcgc cgtgacgagg ggggcgcccg cgggaggcgc caaaacccgg
     1201 cgcggaggcc aggcgcgccc cgatttcgat attcctcgga tctcggatag atcgggattg
     1261 aggatgtagg ttgaacttct ctcgggtgat cccgatctcg gatattcggg attggagtaa
     1321 tacctggcga taggcgctgc caagcaatag ccaaggcgca aggccggcca agagacttgc
     1381 gatagttcgt tcgagaggta gcctgcaaga tggatcggac gaaaggtttg aagattttcc
     1441 gacgagggcc ttggagagtt cggaaattta aatgcaacta tggatgaacg aaatagacag
     1501 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1561 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1621 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1681 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1741 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1801 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1861 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1921 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1981 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2041 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2101 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2161 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2221 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2281 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2341 tggccttttg ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa
     2401 ggaccagatc gaacttgact gacgcaattc tgagcgtagt cgaaacccag acaagtcgat
     2461 ctcgccaagt agctgacttg actggttaat taagtttcga tagcccaagg taaccaatga
     2521 gattagggag ggaagaaagt ttgatgaact acttctgacc cctagtggtg tccagaaaag
     2581 accattaaag gaatgtcttg atcgaaatcg agagatcgcc tgcaggatag ctgacgacct
     2641 taaggataaa tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta
     2701 tccgttgctg aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt
     2761 ttttctgcgc caatgcacct agaatcggtt accaatgctt aatcagtgag gcacctatct
     2821 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2881 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2941 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3001 gtcctgcaac tttatccgcc tccatccagt ctatcaactg ttgccgggaa gctagagtaa
     3061 gtagttcgcc agttaatagt ttacgcaagg ttgttgccat tgctacaggc atcgtggtgt
     3121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctcca atcgttgtca
     3241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga atactcaacc aagtcattct
     3361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3421 cgccacatag cagaacttta aaagtgctca tcattggaaa gcgttcttcg gggcgaaaac
     3481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgc gcacccaact
     3541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatg cgaaagcctc
     3661 ctgctgatcc tgtctcttga tagatgtttg caaaagctag gactcgatta tacgttgacc
     3721 gatcagacta tcaatatcta ggtttaaaca gatcggtacg ctgccgtgac caggcttgag
     3781 tacaatatct gacgattgga cttcctgggt cccaagctgg attccgactg gctgactgcc
     3841 aagcaacgat atggatcgtc gagactcgca ctgagaggat cgttctcggt acaccaggcg
     3901 cgcctagcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     3961 attatctaag actaggctta catactagcc taggcaccgg gcttgcgggt catgcaccag
     4021 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     4081 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcg
     4141 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     4201 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggccc
     4261 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     4321 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     4381 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     4441 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     4501 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     4561 aggcgcaccg tgggcttgta ctcggtcatg tt
//
