LOCUS       sequence-297301-        6645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6645)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6645)
  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-m4252923
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             47..67
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             68..109
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    178..299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(189..208)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     243..262
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      414..460
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      490..626
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    671..742
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(968..1556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1048..1067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2056..3265
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3102..3121
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3269..4297
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4324..4372
                     /note="synthetic polyadenylation signal"
     CDS             join(4638..4706,4707..5498)
                     /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(4856..4875)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5511..5647
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5799..5870
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5875..5921)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5939..5987
                     /note="synthetic polyadenylation signal"
     enhancer        6068..6371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6373..6575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6526..6546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgggg aaacctatac ccaatccact tctcggctta gattctacct aaggatccgc
      121 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
      181 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      241 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      301 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
      361 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
      421 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
      481 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
      541 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
      601 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
      661 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
      721 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
      781 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
      841 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
      901 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1381 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     1801 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     1861 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     1921 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     1981 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     2041 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     2101 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     2161 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     2221 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     2281 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     2341 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     2401 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     2461 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     2521 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     2581 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     2641 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     2701 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     2761 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     2821 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     2881 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca
     2941 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     3001 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     3061 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     3121 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     3181 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     3241 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     3301 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3361 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3421 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     3481 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     3541 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3601 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     3661 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3721 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3781 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     3841 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     3901 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     3961 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4021 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     4081 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4141 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4201 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4261 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     4321 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4381 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     4441 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     4501 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     4561 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     4621 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4681 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4741 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4801 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     4861 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4921 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     4981 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5041 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5101 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5161 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5221 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5281 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5341 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5401 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5461 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     5521 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     5581 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     5641 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     5701 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     5761 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     5821 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     5881 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     5941 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     6001 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6061 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6361 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     6421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     6541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     6601 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
