LOCUS       sequence-297336-        4296 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 4296)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4296)
  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-m4133196
FEATURES             Location/Qualifiers
     source          1..4296
                     /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"
     CDS             join(2289..2357,2358..3149)
                     /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(2507..2526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3162..3298
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3450..3521
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3526..3572)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3590..3638
                     /note="synthetic polyadenylation signal"
     enhancer        3719..4022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4024..4226
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4177..4197
                     /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 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2101 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     2161 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     2221 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     2281 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2341 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2401 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2461 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2521 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2581 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2641 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2701 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2761 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2821 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     2881 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     2941 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3001 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3061 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3121 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     3181 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     3241 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     3301 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     3361 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     3421 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     3481 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     3541 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     3601 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     3661 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     3721 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3781 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3841 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3901 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     3961 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4021 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4081 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4141 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4201 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     4261 tcctaccatc cactcgacac acccgccagc ggccgc
//
