LOCUS       sequence-300270-        6548 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 6548)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6548)
  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-m4252816
FEATURES             Location/Qualifiers
     source          1..6548
                     /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             416..433
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    510..631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(521..540)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     575..594
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      746..792
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      822..958
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1003..1074
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1300..1888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1380..1399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2388..3597
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3434..3453
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3601..4200
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3601..3620)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4097..4117
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4227..4275
                     /note="synthetic polyadenylation signal"
     CDS             join(4541..4609,4610..5401)
                     /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(4759..4778)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5414..5550
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5702..5773
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5778..5824)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5842..5890
                     /note="synthetic polyadenylation signal"
     enhancer        5971..6274
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6276..6478
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6429..6449
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgatg agtgacaaaa tcattcatct aaccgatgac tcctttgata cagacgtgtt
      121 gaaggccgat ggtgcgatac tggtagattt ctgggctgaa tggtgtggcc cgtgcaagat
      181 gatcgcaccc attctggacg agatcgccga cgagtatcaa ggaaaattaa ctgtcgctaa
      241 gcttaatata gatcagaacc cagggacggc acctaaatac ggcatcaggg gaatacctac
      301 cctcctgctc tttaagaatg gggaagttgc cgcgacaaaa gtgggagccc tgtctaaggg
      361 ccagctaaaa gagttcttgg acgctaacct ggcaggttca ggctcggggc acatgcacca
      421 tcaccatcac catagcagcg ggctcgtgcc acgcggctaa gctagcttga ctgactgaga
      481 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      541 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      601 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      661 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
      721 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
      781 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
      841 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
      901 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
      961 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1021 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1081 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1141 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1201 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1741 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2101 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2161 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2221 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2281 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2341 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg
     2401 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     2461 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     2521 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     2581 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     2641 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     2701 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     2761 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     2821 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     2881 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     2941 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     3001 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     3061 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     3121 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     3181 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     3241 ctgttggctt ataatgcagg gtggggccac cagccggtag gtgtgcggta ggcttttctc
     3301 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     3361 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     3421 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     3481 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     3541 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     3601 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     3661 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     3721 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     3781 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     3841 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     3901 tcccggctgg ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag
     3961 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4021 agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg
     4081 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     4141 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag
     4201 gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     4261 ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt
     4321 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     4381 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     4441 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     4501 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     4561 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4621 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     4681 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     4741 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4801 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     4861 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4921 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     4981 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5041 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5101 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     5161 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5221 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5281 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5341 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5401 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     5461 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     5521 gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc
     5581 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     5641 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
     5701 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
     5761 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     5821 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     5881 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
     5941 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     6001 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     6061 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     6121 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6181 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6241 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
     6301 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     6361 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     6421 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     6481 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
     6541 gcggccgc
//
