LOCUS       sequence-297420-        6185 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 6185)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6185)
  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-m4252863
FEATURES             Location/Qualifiers
     source          1..6185
                     /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..94
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    147..268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(158..177)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     212..231
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      383..429
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      459..595
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    640..711
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(937..1525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1017..1036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2025..3234
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3071..3090
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3238..3837
                     /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(3238..3257)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3734..3754
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3864..3912
                     /note="synthetic polyadenylation signal"
     CDS             join(4178..4246,4247..5038)
                     /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(4396..4415)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5051..5187
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5339..5410
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5415..5461)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5479..5527
                     /note="synthetic polyadenylation signal"
     enhancer        5608..5911
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5913..6115
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6066..6086
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgtac ccctatgacg taccagatta tgcgtaagct agcttgactg actgagatac
      121 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
      181 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      241 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
      301 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
      361 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
      421 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
      481 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
      541 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
      601 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
      661 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
      721 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
      781 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
      841 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
      901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1741 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     1801 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     1861 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     1921 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     1981 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt
     2041 tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg
     2101 gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa
     2161 gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac
     2221 tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc
     2281 ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg
     2341 ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg
     2401 gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg
     2461 ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg
     2521 gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg
     2581 ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt
     2641 ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc
     2701 tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact
     2761 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
     2821 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
     2881 ttggcttata atgcagggtg gggccaccag ccggtaggtg tgcggtaggc ttttctccgt
     2941 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     3001 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     3061 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     3121 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     3181 tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg
     3241 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     3301 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     3361 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     3421 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     3481 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     3541 cggctggccg cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc caaggagccc
     3601 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     3661 gccgtcgtgc tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag
     3721 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     3781 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct
     3841 agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg
     3901 gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg
     3961 agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc
     4021 aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt
     4081 taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa
     4141 catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt
     4201 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4261 ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga
     4321 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag
     4381 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4441 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga
     4501 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4561 tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc
     4621 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4681 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt
     4741 gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg
     4801 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4861 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4921 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4981 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc
     5041 gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     5101 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     5161 tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct
     5221 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     5281 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     5341 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     5401 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     5461 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     5521 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     5581 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     5641 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     5701 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     5761 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     5821 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5881 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     5941 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     6001 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     6061 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     6121 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     6181 gccgc
//
