LOCUS       sequence-300619-        4314 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 4314)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4314)
  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-m4133755
FEATURES             Location/Qualifiers
     source          1..4314
                     /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             65..130
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    183..304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(194..213)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     248..267
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      419..465
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      495..631
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    676..747
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(973..1561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1053..1072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2307..2375,2376..3167)
                     /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(2525..2544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3180..3316
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3468..3539
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3544..3590)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3608..3656
                     /note="synthetic polyadenylation signal"
     enhancer        3737..4040
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4042..4244
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4195..4215
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4313..4314,1..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 gatggattat aaggaccacg atggagacta caaagatcat gacattgatt acaaggacga
      121 cgatgacaaa taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
      181 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      241 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      301 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
      361 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
      421 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
      481 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
      541 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
      601 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
      661 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
      721 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
      781 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
      841 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
      901 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1381 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     1801 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     1861 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     1921 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     1981 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2041 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     2101 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     2161 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     2221 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     2281 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     2341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2401 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     2461 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     2521 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2581 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     2641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2701 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     2761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2821 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     2881 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     2941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3001 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     3181 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     3241 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     3301 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     3361 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     3421 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     3481 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     3541 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     3601 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     3661 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     3721 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     3781 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     3841 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     3901 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     3961 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4021 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     4081 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4141 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4201 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     4261 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
