LOCUS       sequence-297271-        6197 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 6197)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6197)
  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-m4252794
FEATURES             Location/Qualifiers
     source          1..6197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..61
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             62..106
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    159..280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(170..189)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     224..243
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      395..441
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      471..607
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    652..723
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(949..1537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1029..1048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2037..3246
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3083..3102
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3250..3849
                     /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(3250..3269)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3746..3766
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3876..3924
                     /note="synthetic polyadenylation signal"
     CDS             join(4190..4258,4259..5050)
                     /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(4408..4427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5063..5199
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5351..5422
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5427..5473)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5491..5539
                     /note="synthetic polyadenylation signal"
     enhancer        5620..5923
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5925..6127
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6078..6098
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6196..6197,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gaaagaaacc gcggcagcta agttcgagag gcagcacatg gacagttaag ctagcttgac
      121 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      181 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      241 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      301 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      361 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      421 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      481 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      541 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      601 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      661 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      721 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      781 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      841 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
      901 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      961 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1021 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1081 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1141 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1201 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1261 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1321 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1381 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1441 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1741 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1801 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1861 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     1921 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     1981 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2041 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2101 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2161 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2221 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2281 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2341 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2401 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2461 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2521 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2581 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     2641 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     2701 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     2761 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     2821 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     2881 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     2941 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3001 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3061 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3121 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3181 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3241 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3301 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3361 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3421 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3481 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3541 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3601 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3661 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     3721 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3781 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     3841 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     3901 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     3961 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4021 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4081 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4141 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4201 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4261 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4321 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4381 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4441 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4501 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4561 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4621 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4681 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4741 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4801 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4861 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4921 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4981 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5041 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5101 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5161 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5221 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5281 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     5341 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     5401 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     5461 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     5521 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5581 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5641 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5701 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5761 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5821 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     5881 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     5941 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6001 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6061 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6121 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6181 cacccgccag cggccgc
//
