LOCUS       sequence-297259-        6194 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 6194)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6194)
  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-m4252793
FEATURES             Location/Qualifiers
     source          1..6194
                     /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..103
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    156..277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(167..186)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     221..240
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      392..438
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      468..604
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    649..720
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(946..1534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1026..1045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2034..3243
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3080..3099
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3247..3846
                     /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(3247..3266)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3743..3763
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3873..3921
                     /note="synthetic polyadenylation signal"
     CDS             join(4187..4255,4256..5047)
                     /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(4405..4424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5060..5196
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5348..5419
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5424..5470)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5488..5536
                     /note="synthetic polyadenylation signal"
     enhancer        5617..5920
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5922..6124
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6075..6095
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6193..6194,1..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 ggggaaacct atacccaatc cacttctcgg cttagattct acctaagcta gcttgactga
      121 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
      181 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      241 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      301 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      361 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
      421 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
      481 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
      541 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
      601 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
      661 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
      721 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
      781 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
      841 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
      901 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      961 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1021 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1081 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1141 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1201 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1261 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1321 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1381 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1441 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1501 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1561 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1621 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1681 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1741 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     1801 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     1861 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     1921 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     1981 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     2041 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     2101 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     2161 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     2221 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     2281 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     2341 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     2401 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     2461 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     2521 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     2581 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     2641 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     2701 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     2761 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     2821 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     2881 cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt gcggtaggct
     2941 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     3001 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     3061 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     3121 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     3181 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     3241 gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3301 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     3361 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     3421 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     3481 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     3541 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
     3601 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     3661 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
     3721 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     3781 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     3841 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     3901 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     3961 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     4021 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     4081 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     4141 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     4201 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4261 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     4321 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     4381 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4441 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     4501 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4561 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     4621 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4681 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4741 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     4801 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4861 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4921 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4981 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5041 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     5101 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     5161 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     5221 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     5281 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     5341 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     5401 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     5461 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     5521 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     5581 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5641 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5701 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5761 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     5821 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     5881 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     5941 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     6001 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     6061 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     6121 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     6181 ccgccagcgg ccgc
//
