LOCUS       sequence-297933-        6557 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 6557)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6557)
  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-m4133373
FEATURES             Location/Qualifiers
     source          1..6557
                     /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..466
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    519..640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(530..549)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     584..603
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      755..801
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      831..967
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1012..1083
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1309..1897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1389..1408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2397..3606
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3443..3462
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3610..4209
                     /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(3610..3629)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4106..4126
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4236..4284
                     /note="synthetic polyadenylation signal"
     CDS             join(4550..4618,4619..5410)
                     /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(4768..4787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5423..5559
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5711..5782
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5787..5833)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5851..5899
                     /note="synthetic polyadenylation signal"
     enhancer        5980..6283
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6285..6487
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6438..6458
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6556..6557,1..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 tttcagtcgt acccctatga cgtaccagat tatgcgtaag ctagcttgac
      481 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      541 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      601 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      661 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      721 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      781 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      841 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      901 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      961 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     1021 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     1081 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1141 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1201 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2101 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     2161 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2221 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2281 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2341 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2401 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2461 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2521 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2581 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2641 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2701 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2761 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2821 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2881 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2941 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     3001 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     3061 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     3121 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     3181 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     3241 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     3301 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3361 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3421 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3481 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3541 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3601 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3661 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3721 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3781 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3841 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3901 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3961 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     4021 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     4081 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     4141 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4201 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     4261 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     4321 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4381 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4441 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4501 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4561 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4621 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4681 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4741 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4801 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4861 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4921 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4981 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5041 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5101 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5161 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5221 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5281 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5341 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5401 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5461 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5521 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5581 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5641 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     5701 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     5761 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     5821 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     5881 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5941 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6001 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6061 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6121 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6181 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6241 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     6301 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6361 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6421 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6481 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6541 cacccgccag cggccgc
//
