LOCUS       sequence-300754-        6630 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 6630)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6630)
  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-m4252960
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        121..424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    505..553
                     /note="synthetic polyadenylation signal"
     terminator      571..617
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    622..693
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(845..981)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(994..1785,1786..1854))
                     /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     1617..1636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2120..2168
                     /note="synthetic polyadenylation signal"
     CDS             complement(2195..3223)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3227..4436)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3371..3390)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4936..5524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5425..5444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5750..5821
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5866..6002)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6032..6078)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6193..6314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6230..6249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6284..6303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6383..6409)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(6410..6430)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(join(6547..6630,1..119))
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6576..6596)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
       61 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcag
      121 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
      181 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
      241 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
      301 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
      361 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
      421 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actagatctg
      481 gctacactcg atcagcagtt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
      541 ggttttttgt gtgtgtaacg caacgattga tagcataacc ccttggggcc tctaaacggg
      601 tcttgagggg ttttttgaat tgagaggtag ctgaagctgc tgttatgacc actagaggga
      661 agaagatacc tgtggagcta atggtccaag atgtcgggag ccgcgatcgc tgccaaaaag
      721 gtaagctgac gatagctgac gagattcgat gcccgacgag caaatgagcg ccacggctgt
      781 tcgcgtaagc tcgaaacgac ggagattcga tagagccaga gtcgagttta aacgattcgg
      841 gatcttaagg ataaatttct ggtaaggagg acacgtatgg aagtgggcaa gttggggaag
      901 ccgtatccgt tgctgaatct ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg
      961 catttttttc tgcgccaatg cacctagaat cggttaccaa tgcttaatca gtgaggcacc
     1021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     1141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1201 aagtggtcct gcaactttat ccgcctccat ccagtctatc aactgttgcc gggaagctag
     1261 agtaagtagt tcgccagtta atagtttacg caaggttgtt gccattgcta caggcatcgt
     1321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccaatcgt
     1441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgaatact caaccaagtc
     1561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaagcgtt cttcggggcg
     1681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgcgcacc
     1741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatgcgaaa
     1861 gcctcctgct gatcctgtct cttgatagat gtttgcaaaa gctaggactc gattatacgt
     1921 tgaccgatca gactatcaat atctaggttt aaacagatcg gtacgctgcc gtgaccaggc
     1981 ttgagtacaa tatctgacga ttggacttcc tgggtcccaa gctggattcc gactggctga
     2041 ctgccaagca acgatatgga tcgtcgagac tcgcactgag aggatcgttc tcggtacacc
     2101 aggcgcgcct agcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg
     2161 attttattat ctaagactag gcttacatac tagcttattc ctttgccctc ggacgagtgc
     2221 tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggcgg
     2281 cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt
     2341 cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga
     2401 gttggtcaag accaatgcgg agcatatacg cccggagccg gggcgatcct gcaagctccg
     2461 gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga
     2521 agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga
     2581 ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga
     2641 ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga
     2701 cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa
     2761 tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc
     2821 cgaacccgct cgtctggcta agatcggcgg cagcgatggc atccattgcc tccgcgaccg
     2881 gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac
     2941 ggcgggagat gcaataggtc aggctctcgc tgaactcccc aatgtcaagc acttccggaa
     3001 tcgggagcgc ggcggatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg
     3061 cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag
     3121 attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa
     3181 acttctcgac agacgtcgcg gtgagttcag gctttttacc catgttgtct aacaaaaaag
     3241 ccaaaaacgg ccagaattta gcggacaatt tacaagtcta acactgaaaa ttacatattg
     3301 acccaaatga ttacatttca aaaggtgcct aaaaaacttc acaaaacaca ctcgccaacc
     3361 ccgagcgcat agttcaaaac cggagcttca gctacttaag aagataggta cataaaaccg
     3421 accaaagaaa ctgacgcctc acttatccct cccctcacca gaggtccggc gcctgtcgat
     3481 tcaggagagc ctaccctagg cccgaaccct gcgtcctgcg acggagaaaa gcctaccgca
     3541 cacctaccgg ctggtggccc caccctgcat tataagccaa cagaacgggt gacgtcacga
     3601 cacgacgagg gcgcgcgctc ccaaaggtac gggtgcactg cccaacggca ccgccataac
     3661 tgccgccccc gcaacagacg acaaaccgag ttctccagtc agtgacaaac ttcacgtcag
     3721 ggtccccaga tggtgcccag cccatctcac ccgaataaga gctttcccgc attagcgaag
     3781 ggctcaagac cttgggttct tgccgcccac catgcccccc accttgtttc aacgacctca
     3841 cagcccgcct cacaagcgtc ttccattcaa gactcgggaa cagccgccat tttgctgcgc
     3901 tccccccaac ccccagttca gggcaacctt gctcgcggac ccagactaca gcccttggcg
     3961 gtctctccac acgcttccgt cccaccgagc ggcccggcgg ccacgaaagc cccggccagc
     4021 ccagcagccc gctactcacc aagtgacgat cacagcgatc cacaaacaag aaccgcgacc
     4081 caaatcccgg ctgcgacgga actagctgtg ccacacccgg cgcgtcctta tataatcatc
     4141 ggcgttcacc gccccacgga gatccctccg cagaatcgcc gagaagggac tacttttcct
     4201 cgcctgttcc gctctctgga aagaaaacca gtgccctaga gtcacccaag tcccgtccta
     4261 aaatgtcctt ctgctgatac tggggttcta aggccgagtc ttatgagcag cgggccgctg
     4321 tcctgagcgt ccgggcggaa ggatcaggac gctcctgcgc ccttcgtctg acgtggcagc
     4381 gctcgccgtg acgagggggg cgcccgcggg aggcgccaaa acccggcgcg gaggccaggc
     4441 gcgccccgat ttcgatattc ctcggatctc ggatagatcg ggattgagga tgtaggttga
     4501 acttctctcg ggtgatcccg atctcggata ttcgggattg gagtaatacc tggcgatagg
     4561 cgctgccaag caatagccaa ggcgcaaggc cggccaagag acttgcgata gttcgttcga
     4621 gaggtagcct gcaagatgga tcggacgaaa ggtttgaaga ttttccgacg agggccttgg
     4681 agagttcgga aatttaaatg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4741 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4801 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4861 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4921 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4981 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5041 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5101 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5161 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5221 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5281 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5341 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5401 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5461 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5521 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5581 catgatttaa atgaaactcg taatcggaga acgggtgcaa tagcaaggac cagatcgaac
     5641 ttgactgacg caattctgag cgtagtcgaa acccagacaa gtcgatctcg ccaagtagct
     5701 gacttgactg gttaattaag tttcgatagc ccaaggtaac caatgagatt agggagggaa
     5761 gaaagtttga tgaactactt ctgaccccta gtggtgtcca gaaaagacca ttaaaggaat
     5821 gtcttgatcg aaatcgagag atcgcctgca ggatagctga cgaccttaag gataaatttc
     5881 tggtaaggag gacacgtatg gaagtgggca agttggggaa gccgtatccg ttgctgaatc
     5941 tggcatatgt gggagtataa gacgcgcagc gtcgcatcag gcattttttt ctgcgccaat
     6001 gccggtagat agcaatccgg cccgaggggc acaaaaaacc cctcaagacc cgtttagagg
     6061 ccccaagggg ttatgctacg ataccgatag agatgggcca ataccacatt tgtagaggtt
     6121 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     6181 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6241 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6301 atcaatgtat cttatcatgt ctgtgagctg aaggtacgct gtatctcagt cagtcaagct
     6361 agcagaggcc tgcggatcct tacgcataat ctggtacgtc ataggggtac gactgaaagt
     6421 aaaggttttc cactgcgaaa ccgttgagaa aggactgagc atcggcactc atggtggcgg
     6481 ccgctggcgg gtgtgtcgag tggatggtag gatcgacaaa gatctgatct gacggttcac
     6541 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     6601 tggggcggag ttgttacgac attttggaaa
//
