LOCUS       sequence-307268-        4650 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 4650)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4650)
  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-m4134050
FEATURES             Location/Qualifiers
     source          1..4650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      657..666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1077..1097
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1098..1124
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1177..1298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1188..1207)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1242..1261
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1413..1459
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1489..1625
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1670..1741
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1967..2555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3301..3369,3370..4161)
                     /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(3519..3538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4174..4310
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4462..4533
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4538..4584)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4602..4650
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca
      661 ccatgggtaa tgcaacagaa gtgcctgcca attcaactgt attatctttc tgtgcttttg
      721 ctgtagatgc tgctaaagct tacaaagatt atctagctag tgggggacaa ccaatcacta
      781 attgtgttaa gatgttgtgt acacacactg gtactggtca ggcaataaca gttacaccgg
      841 aagccaatat ggatcaagaa tcctttggtg gtgcatcgtg ttgtctgtac tgccgttgcc
      901 acatagatca tccaaatcct aaaggatttt gtgacttaaa aggtaagtat gtacaaatac
      961 ctacaacttg tgctaatgac cctgtgggtt ttacacttaa aaacacagtc tgtaccgtct
     1021 gcggtatgtg gaaaggttat ggctgtagtt gtgatcaact ccgcgaaccc atgcttgaaa
     1081 acctttactt tcagtcgtac ccctatgacg taccagatta tgcgtaagct agcttgactg
     1141 actgagatac agcgtacctt cagctcacag acatgataag atacattgat gagtttggac
     1201 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1261 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1321 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1381 aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg
     1441 ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag
     1501 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     1561 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     1621 cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa
     1681 tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct
     1741 aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg
     1801 cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg
     1861 gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag
     1921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2401 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2761 cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct
     2821 cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac
     2881 tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca
     2941 ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca
     3001 tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta
     3061 ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc
     3121 agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg
     3181 tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat
     3241 aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc
     3301 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3361 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg
     3421 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3481 gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3541 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3601 gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3661 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt
     3721 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3781 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3841 cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct
     3901 tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3961 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4021 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4081 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4141 tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat
     4201 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     4261 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat
     4321 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     4381 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     4441 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     4501 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     4561 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     4621 catccatctt cgatggatag cgattttatt
//
