LOCUS       sequence-300732-        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"
     misc_feature    151..222
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(227..273)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    291..339
                     /note="synthetic polyadenylation signal"
     enhancer        420..723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        725..927
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     878..898
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1044..1064
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1065..1091
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1144..1265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1155..1174)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1209..1228
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1380..1426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1456..1592
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1637..1708
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1934..2522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2014..2033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3022..4231
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4068..4087
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4235..4834
                     /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(4235..4254)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4731..4751
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4861..4909
                     /note="synthetic polyadenylation signal"
     CDS             join(5175..5243,5244..6035)
                     /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(5393..5412)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6048..6184
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
       61 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
      121 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
      181 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
      241 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
      301 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
      361 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      421 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      481 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      541 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      601 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      661 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      721 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      781 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      841 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      901 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg
      961 atcctaccat ccactcgaca cacccgccag cggccgccac catgagtgcc gatgctcagt
     1021 cctttctcaa cggtttcgca gtggaaaacc tttactttca gtcgtacccc tatgacgtac
     1081 cagattatgc gtaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
     1141 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1201 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1261 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1321 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
     1381 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
     1441 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     1501 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     1561 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
     1621 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
     1681 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     1741 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     1801 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     1861 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2101 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2341 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     2761 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     2821 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     2881 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     2941 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     3001 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     3061 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     3121 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     3181 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     3241 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     3301 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     3361 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     3421 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     3481 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     3541 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     3601 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     3661 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     3721 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     3781 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     3841 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     3901 ccaccagccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     3961 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     4021 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     4081 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     4141 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     4201 attctggccg tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg
     4261 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4321 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4381 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4441 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4501 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4561 ggagggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4621 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4681 ggccgccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4741 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4801 cacctggtgc atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat
     4861 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     4921 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     4981 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     5041 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     5101 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     5161 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5221 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5281 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5341 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5401 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5461 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     5521 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5581 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5641 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5701 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5761 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     5821 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5881 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5941 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6001 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     6061 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     6121 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     6181 ttaag
//
