LOCUS       sequence-369842-        4592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp6.
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-m4134006
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      1584..2172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2073..2092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2398..2469
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2514..2650)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2663..3454,3455..3523))
                     /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     3286..3305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    3789..3837
                     /note="synthetic polyadenylation signal"
     CDS             complement(3864..4463)
                     /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(3947..3967)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4444..4463
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(4467..4592,1..1084))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 cgccaacccc gagcgcatag ttcaaaaccg gagcttcagc tacttaagaa gataggtaca
       61 taaaaccgac caaagaaact gacgcctcac ttatccctcc cctcaccaga ggtccggcgc
      121 ctgtcgattc aggagagcct accctaggcc cgaaccctgc gtcctgcgac ggagaaaagc
      181 ctaccgcaca cctaccggca ggtggcccca ccctgcatta taagccaaca gaacgggtga
      241 cgtcacgaca cgacgagggc gcgcgctccc aaaggtacgg gtgcactgcc caacggcacc
      301 gccataactg ccgcccccgc aacagacgac aaaccgagtt ctccagtcag tgacaaactt
      361 cacgtcaggg tccccagatg gtgcccagcc catctcaccc gaataagagc tttcccgcat
      421 tagcgaaggg ctcaagacct tgggttcttg ccgcccacca tgccccccac cttgtttcaa
      481 cgacctcaca gcccgcctca caagcgtctt ccattcaaga ctcgggaaca gccgccattt
      541 tgctgcgctc cccccaaccc ccagttcagg gcaaccttgc tcgcggaccc agactacagc
      601 ccttggcggt ctctccacac gcttccgtcc caccgagcgg cccggcggcc acgaaagccc
      661 cggccagccc agcagcccgc tactcaccaa gtgacgatca cagcgatcca caaacaagaa
      721 ccgcgaccca aatcccggct gcgacggaac tagctgtgcc acacccggcg cgtccttata
      781 taatcatcgg cgttcaccgc cccacggaga tccctccgca gaatcgccga gaagggacta
      841 cttttcctcg cctgttccgc tctctggaaa gaaaaccagt gccctagagt cacccaagtc
      901 ccgtcctaaa atgtccttct gctgatactg gggttctaag gccgagtctt atgagcagcg
      961 ggccgctgtc ctgagcgtcc gggcggaagg atcaggacgc tcctgcgccc ttcgtctgac
     1021 gtggcagcgc tcgccgtgac gaggggggcg cccgcgggag gcgccaaaac ccggcgcgga
     1081 ggccaggcgc gccccgattt cgatattcct cggatctcgg atagatcggg attgaggatg
     1141 taggttgaac ttctctcggg tgatcccgat ctcggatatt cgggattgga gtaatacctg
     1201 gcgataggcg ctgccaagca atagccaagg cgcaaggccg gccaagagac ttgcgatagt
     1261 tcgttcgaga ggtagcctgc aagatggatc ggacgaaagg tttgaagatt ttccgacgag
     1321 ggccttggag agttcggaaa tttaaatgca actatggatg aacgaaatag acagatcgct
     1381 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1441 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1501 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1561 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1621 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1681 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1741 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1801 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1861 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1921 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1981 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2041 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2101 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2161 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2221 tttgctcaca tgatttaaat gaaactcgta atcggagaac gggtgcaata gcaaggacca
     2281 gatcgaactt gactgacgca attctgagcg tagtcgaaac ccagacaagt cgatctcgcc
     2341 aagtagctga cttgactggt taattaagtt tcgatagccc aaggtaacca atgagattag
     2401 ggagggaaga aagtttgatg aactacttct gacccctagt ggtgtccaga aaagaccatt
     2461 aaaggaatgt cttgatcgaa atcgagagat cgcctgcagg atagctgacg accttaagga
     2521 taaatttctg gtaaggagga cacgtatgga agtgggcaag ttggggaagc cgtatccgtt
     2581 gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt cgcatcaggc atttttttct
     2641 gcgccaatgc acctagaatc ggttaccaat gcttaatcag tgaggcacct atctcagcga
     2701 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2761 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2821 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2881 caactttatc cgcctccatc cagtctatca actgttgccg ggaagctaga gtaagtagtt
     2941 cgccagttaa tagtttacgc aaggttgttg ccattgctac aggcatcgtg gtgtcacgct
     3001 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3061 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccaatcgtt gtcagaagta
     3121 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3181 tgccatccgt aagatgcttt tctgtgactg gtgaatactc aaccaagtca ttctgagaat
     3241 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3301 atagcagaac tttaaaagtg ctcatcattg gaaagcgttc ttcggggcga aaactctcaa
     3361 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgcgcaccc aactgatctt
     3421 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3481 caaaaaaggg aataagggcg acacggaaat gttgaatact catgcgaaag cctcctgctg
     3541 atcctgtctc ttgatagatg tttgcaaaag ctaggactcg attatacgtt gaccgatcag
     3601 actatcaata tctaggttta aacagatcgg tacgctgccg tgaccaggct tgagtacaat
     3661 atctgacgat tggacttcct gggtcccaag ctggattccg actggctgac tgccaagcaa
     3721 cgatatggat cgtcgagact cgcactgaga ggatcgttct cggtacacca ggcgcgccta
     3781 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     3841 taagactagg cttacatact agcctaggca ccgggcttgc gggtcatgca ccaggtgcgc
     3901 ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg
     3961 aggttgcggg gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggcggcctcc
     4021 actccgggga gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg
     4081 acggtggcca ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccctccatc
     4141 tgttgctgcg cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg
     4201 aacaccgccc ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg
     4261 tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc
     4321 tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg
     4381 gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc
     4441 accgtgggct tgtactcggt catgttgtct aacaaaaaag ccaaaaacgg ccagaattta
     4501 gcggacaatt tacaagtcta acactgaaaa ttacatattg acccaaatga ttacatttca
     4561 aaaggtgcct aaaaaacttc acaaaacaca ct
//
