LOCUS       sequence-297361-        6621 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 6621)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6621)
  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-m4252963
FEATURES             Location/Qualifiers
     source          1..6621
                     /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..85
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    154..275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(165..184)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     219..238
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      390..436
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      466..602
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    647..718
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(944..1532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1024..1043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2032..3241
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3078..3097
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3245..4273
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4300..4348
                     /note="synthetic polyadenylation signal"
     CDS             join(4614..4682,4683..5474)
                     /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(4832..4851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5487..5623
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5775..5846
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5851..5897)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5915..5963
                     /note="synthetic polyadenylation signal"
     enhancer        6044..6347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6349..6551
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6502..6522
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgcac catcaccatc accattaagg atccgcaggc ctctgctagc ttgactgact
      121 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
      181 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      241 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      301 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
      361 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
      421 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
      481 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
      541 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
      601 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
      661 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
      721 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
      781 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
      841 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
      901 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      961 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1021 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1081 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1141 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1201 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1261 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1321 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1381 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1441 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1501 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1561 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1621 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1681 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1741 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     1801 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     1861 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     1921 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     1981 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     2041 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     2101 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     2161 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     2221 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     2281 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     2341 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     2401 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     2461 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     2521 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     2581 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     2641 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     2701 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     2761 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     2821 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     2881 cgttctgttg gcttataatg cagggtgggg ccaccagccg gtaggtgtgc ggtaggcttt
     2941 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     3001 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     3061 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     3121 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     3181 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     3241 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     3301 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     3361 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     3421 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     3481 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     3541 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     3601 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     3661 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     3721 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     3781 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     3841 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     3901 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     3961 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4021 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4081 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     4141 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     4201 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     4261 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     4321 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     4381 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     4441 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     4501 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     4561 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     4621 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4681 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     4741 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4801 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4861 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4921 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4981 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     5041 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5101 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5161 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     5221 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5281 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5341 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5401 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5461 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     5521 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     5581 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     5641 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     5701 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     5761 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     5821 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     5881 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     5941 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     6001 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     6061 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     6121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     6181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     6241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     6301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     6361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     6421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     6481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     6541 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     6601 gacacacccg ccagcggccg c
//
