LOCUS       sequence-297610-        6131 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 6131)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6131)
  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-m4252976
FEATURES             Location/Qualifiers
     source          1..6131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    93..214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(104..123)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     158..177
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      329..375
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      405..541
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    586..657
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(883..1471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(963..982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1971..3180
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3017..3036
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3184..3783
                     /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(3184..3203)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3680..3700
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3810..3858
                     /note="synthetic polyadenylation signal"
     CDS             join(4124..4192,4193..4984)
                     /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(4342..4361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4997..5133
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5285..5356
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5361..5407)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5425..5473
                     /note="synthetic polyadenylation signal"
     enhancer        5554..5857
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5859..6061
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6012..6032
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6130..6131,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg taagctagct tgactgactg
       61 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
      121 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
      181 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
      241 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
      301 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
      361 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
      421 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
      481 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
      541 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
      601 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
      661 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
      721 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
      781 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
      841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1681 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     1741 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     1801 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     1861 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     1921 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg
     1981 ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga
     2041 cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc
     2101 tcataagact cggccttaga accccagtat cagcagaagg acattttagg acgggacttg
     2161 ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc
     2221 cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag
     2281 gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg
     2341 tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt
     2401 tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta
     2461 gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg
     2521 cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa
     2581 caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg
     2641 gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt
     2701 gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc
     2761 gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc
     2821 gttctgttgg cttataatgc agggtggggc caccagccgg taggtgtgcg gtaggctttt
     2881 ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg
     2941 gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc
     3001 tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg
     3061 ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt
     3121 cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac
     3181 aacatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     3241 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     3301 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     3361 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     3421 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     3481 ggttcccggc tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag
     3541 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     3601 ggcagcgccg tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc
     3661 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     3721 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     3781 taggctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt
     3841 gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc
     3901 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     3961 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     4021 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     4081 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     4141 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4201 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     4261 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     4321 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4381 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     4441 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4501 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     4561 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4621 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4681 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     4741 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4801 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4861 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4921 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4981 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     5041 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     5101 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     5161 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     5221 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     5281 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     5341 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     5401 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     5461 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     5521 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     5581 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     5641 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     5701 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     5761 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     5821 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
     5881 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     5941 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     6001 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     6061 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
     6121 ccagcggccg c
//
