LOCUS       sequence-297443-        6642 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 6642)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6642)
  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-m4252896
FEATURES             Location/Qualifiers
     source          1..6642
                     /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             62..106
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    175..296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(186..205)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     240..259
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      411..457
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      487..623
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    668..739
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(965..1553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1045..1064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2053..3262
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3099..3118
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3266..4294
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4321..4369
                     /note="synthetic polyadenylation signal"
     CDS             join(4635..4703,4704..5495)
                     /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(4853..4872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5508..5644
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5796..5867
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5872..5918)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5936..5984
                     /note="synthetic polyadenylation signal"
     enhancer        6065..6368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6370..6572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6523..6543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6641..6642,1..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 gaaagaaacc gcggcagcta agttcgagag gcagcacatg gacagttaag gatccgcagg
      121 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
      181 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      241 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
      301 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      361 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
      421 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
      481 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
      541 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
      601 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
      661 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
      721 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
      781 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
      841 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
      901 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      961 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1021 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1081 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1141 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1201 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1261 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1321 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1381 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1441 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1501 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1561 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1621 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1681 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1741 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     1801 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     1861 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     1921 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     1981 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     2041 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     2101 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     2161 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2221 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2281 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2341 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2401 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     2461 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     2521 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     2581 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     2641 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     2701 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     2761 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     2821 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     2881 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc
     2941 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     3001 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3061 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3121 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3181 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3241 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     3301 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3361 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3421 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     3481 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     3541 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     3601 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     3661 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     3721 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     3781 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     3841 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     3901 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     3961 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4021 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     4081 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4141 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4201 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4261 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     4321 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     4381 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     4441 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     4501 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     4561 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     4621 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4681 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4741 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4801 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4861 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4921 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     4981 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5041 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5101 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5161 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5221 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     5281 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5341 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5401 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5461 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     5521 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     5581 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     5641 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     5701 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     5761 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     5821 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     5881 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     5941 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     6001 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     6061 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6361 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     6421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     6481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     6541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     6601 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
