LOCUS       sequence-295408-        6565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6565)
  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-m4133182
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             419..439
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             440..481
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    534..655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(545..564)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     599..618
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      770..816
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      846..982
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1027..1098
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1324..1912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1404..1423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2412..3621
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3458..3477
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3625..4224
                     /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(3625..3644)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4121..4141
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4251..4299
                     /note="synthetic polyadenylation signal"
     CDS             join(4565..4633,4634..5425)
                     /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(4783..4802)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5438..5574
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5719..5790
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5795..5841)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5859..5907
                     /note="synthetic polyadenylation signal"
     enhancer        5988..6291
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6293..6495
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6446..6466
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6564..6565,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggt aatgcaacag aagtgcctgc caattcaact gtattatctt tctgtgcttt
       61 tgctgtagat gctgctaaag cttacaaaga ttatctagct agtgggggac aaccaatcac
      121 taattgtgtt aagatgttgt gtacacacac tggtactggt caggcaataa cagttacacc
      181 ggaagccaat atggatcaag aatcctttgg tggtgcatcg tgttgtctgt actgccgttg
      241 ccacatagat catccaaatc ctaaaggatt ttgtgactta aaaggtaagt atgtacaaat
      301 acctacaact tgtgctaatg accctgtggg ttttacactt aaaaacacag tctgtaccgt
      361 ctgcggtatg tggaaaggtt atggctgtag ttgtgatcaa ctccgcgaac ccatgcttga
      421 aaacctttac tttcagtcgg ggaaacctat acccaatcca cttctcggct tagattctac
      481 ctaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
      541 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      601 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      661 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
      721 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
      781 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
      841 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
      901 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
      961 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     1021 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     1081 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     1141 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     1201 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     1261 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1321 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1381 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1441 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1501 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1561 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1621 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1681 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1741 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1801 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1861 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1921 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1981 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2041 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2101 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     2161 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     2221 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     2281 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     2341 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     2401 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac
     2461 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     2521 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     2581 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     2641 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     2701 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     2761 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     2821 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     2881 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     2941 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     3001 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     3061 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct
     3121 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     3181 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     3241 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     3301 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     3361 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     3421 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     3481 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     3541 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg
     3601 tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc
     3661 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     3721 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     3781 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     3841 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     3901 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc
     3961 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4021 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag
     4081 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4141 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4201 atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg
     4261 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg
     4321 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc
     4381 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc
     4441 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac
     4501 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt
     4561 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4621 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4681 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4741 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4801 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4861 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4921 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4981 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5041 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5101 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct
     5161 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct
     5221 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5281 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5341 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5401 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc
     5461 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     5521 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatcgt
     5581 ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg
     5641 gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc
     5701 agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag
     5761 tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag
     5821 aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat
     5881 ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag
     5941 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     6001 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     6061 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     6121 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     6181 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     6241 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg
     6301 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     6361 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     6421 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     6481 tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc
     6541 actcgacaca cccgccagcg gccgc
//
