LOCUS       sequence-297300-        6672 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 6672)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6672)
  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-m4252908
FEATURES             Location/Qualifiers
     source          1..6672
                     /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             71..136
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    205..326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(216..235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     270..289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      441..487
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      517..653
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    698..769
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(995..1583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1075..1094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2083..3292
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3129..3148
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3296..4324
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4351..4399
                     /note="synthetic polyadenylation signal"
     CDS             join(4665..4733,4734..5525)
                     /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(4883..4902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5538..5674
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5826..5897
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5902..5948)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5966..6014
                     /note="synthetic polyadenylation signal"
     enhancer        6095..6398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6400..6602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6553..6573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgatg gattataagg accacgatgg agactacaaa gatcatgaca ttgattacaa
      121 ggacgacgat gacaaataag gatccgcagg cctctgctag cttgactgac tgagatacag
      181 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
      241 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      301 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      361 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
      421 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
      481 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
      541 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
      601 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
      661 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
      721 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
      781 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
      841 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
      901 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
      961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1501 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1561 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1621 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1801 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     1861 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     1921 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     1981 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     2041 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     2101 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     2161 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2221 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2281 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2341 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2401 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     2461 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     2521 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     2581 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     2641 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     2701 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     2761 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     2821 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     2881 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     2941 ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg
     3001 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     3061 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     3121 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     3181 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     3241 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     3301 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3361 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3421 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3481 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     3541 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     3601 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     3661 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     3721 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     3781 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     3841 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     3901 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     3961 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4021 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     4081 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4141 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4201 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     4261 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4321 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     4381 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     4441 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     4501 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     4561 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     4621 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     4681 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4741 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     4801 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     4861 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4921 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     4981 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5041 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5101 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5161 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5221 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     5281 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5341 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5401 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5461 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5521 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     5581 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     5641 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     5701 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     5761 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     5821 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     5881 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     5941 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     6001 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     6061 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6361 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     6421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     6601 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     6661 gccagcggcc gc
//
