LOCUS       sequence-295222-        5019 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 5019)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5019)
  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-m4133873
FEATURES             Location/Qualifiers
     source          1..5019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..445
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             794..811
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    888..1009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(899..918)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     953..972
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1124..1170
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1200..1336
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1381..1452
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1678..2266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1758..1777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3012..3080,3081..3872)
                     /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(3230..3249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3885..4021
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4173..4244
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4249..4295)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4313..4361
                     /note="synthetic polyadenylation signal"
     enhancer        4442..4745
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4747..4949
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4900..4920
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5018..5019,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gggaatgcga cagaggttcc tgccaactca acggtcttgt ctttttgtgc
       61 tttcgccgtg gacgctgcaa aagcctacaa ggattatcta gcctcgggtg gacagccaat
      121 cactaactgc gtaaagatgc tttgtaccca caccggcact ggccaagcaa ttacagtgac
      181 ccccgaagct aatatggacc aggagtcctt cggaggtgcg agctgctgcc tgtactgtcg
      241 ttgtcatata gatcacccga atcccaaagg gttttgcgac ctcaagggca aatatgtcca
      301 gatcccaacg acatgcgcca acgatcctgt gggattcact ctgaagaaca ccgtatgtac
      361 cgtttgcggg atgtggaagg ggtatggctg tagttgcgac caattacggg aaccgatgct
      421 gcaagaaaac ctttactttc agtcgatgag tgacaaaatc attcatctaa ccgatgactc
      481 ctttgataca gacgtgttga aggccgatgg tgcgatactg gtagatttct gggctgaatg
      541 gtgtggcccg tgcaagatga tcgcacccat tctggacgag atcgccgacg agtatcaagg
      601 aaaattaact gtcgctaagc ttaatataga tcagaaccca gggacggcac ctaaatacgg
      661 catcagggga atacctaccc tcctgctctt taagaatggg gaagttgccg cgacaaaagt
      721 gggagccctg tctaagggcc agctaaaaga gttcttggac gctaacctgg caggttcagg
      781 ctcggggcac atgcaccatc accatcacca tagcagcggg ctcgtgccac gcggctaagc
      841 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      901 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      961 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1021 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1081 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1141 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1201 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1261 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1321 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     1381 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1441 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1501 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1561 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2101 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2521 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2581 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2641 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2701 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2761 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2821 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     2881 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     2941 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     3001 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3061 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3121 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3181 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3241 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3301 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3361 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3421 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3481 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3541 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3601 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3661 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3721 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3781 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3841 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     3901 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     3961 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     4021 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     4081 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     4141 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     4201 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     4261 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     4321 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     4381 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4441 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4501 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4561 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4621 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4681 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4741 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4801 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4861 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4921 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     4981 cgatcctacc atccactcga cacacccgcc agcggccgc
//
