LOCUS       sequence-299529-        6203 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 6203)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6203)
  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-m4258295
FEATURES             Location/Qualifiers
     source          1..6203
                     /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..112
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    165..286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(176..195)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     230..249
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      401..447
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      477..613
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    658..729
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(955..1543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1035..1054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2043..3252
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3089..3108
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3256..3855
                     /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(3256..3275)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3752..3772
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3882..3930
                     /note="synthetic polyadenylation signal"
     CDS             join(4196..4264,4265..5056)
                     /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(4414..4433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5069..5205
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5357..5428
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5433..5479)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5497..5545
                     /note="synthetic polyadenylation signal"
     enhancer        5626..5929
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5931..6133
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6084..6104
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgaaa gaaaccgcgg cagctaagtt cgagaggcag cacatggaca gttaagctag
      121 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      181 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      241 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      301 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      361 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
      421 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
      481 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
      541 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
      601 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
      661 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
      721 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
      781 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
      841 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
      901 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      961 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1021 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1081 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1141 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1201 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1261 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1381 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1561 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1621 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1681 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1741 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     1801 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     1861 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     1921 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     1981 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2041 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2101 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2161 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     2221 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     2281 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     2341 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     2401 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     2461 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     2521 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     2581 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     2641 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     2701 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     2761 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     2821 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     2881 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg
     2941 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3001 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3061 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3121 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3181 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     3241 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3301 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3361 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3421 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3481 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3541 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     3601 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     3661 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     3721 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3781 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     3841 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     3901 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     3961 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4021 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4081 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4141 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     4321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4381 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4501 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     4621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4741 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     4801 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5041 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5101 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5161 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     5221 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     5281 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     5341 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     5401 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     5461 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     5521 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     5581 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5641 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5701 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5761 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5821 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5881 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     5941 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6001 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6061 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6121 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     6181 tcgacacacc cgccagcggc cgc
//
