LOCUS       sequence-300487-        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    96..144
                     /note="synthetic polyadenylation signal"
     CDS             complement(171..770)
                     /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(254..274)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     751..770
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(774..1983)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(918..937)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      2483..3071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2972..2991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3297..3368
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3413..3549)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3579..3625)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3740..3861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3777..3796)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3831..3850
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     regulatory      3946..3955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4024..4226)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4053..4073)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4228..4531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4612..4660
                     /note="synthetic polyadenylation signal"
     terminator      4678..4724
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    4729..4800
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4952..5088)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(5101..5892,5893..5961))
                     /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     5724..5743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gattccgact ggctgactgc caagcaacga tatggatcgt cgagactcgc actgagagga
       61 tcgttctcgg tacaccaggc gcgcctagcg ttacacacac aaaaaaccaa cacacatcca
      121 tcttcgatgg atagcgattt tattatctaa gactaggctt acatactagc ctaggcaccg
      181 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg
      241 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg
      301 ggcaccccgg cgcgctcggc ggcctccact ccggggagca cgacggcgct gcccagaccc
      361 ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc
      421 cggtgcggcg ccaggaggcc ctccatctgt tgctgcgcgg ccagccggga accgctcaac
      481 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg
      541 gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg
      601 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg
      661 acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg
      721 gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat gttgtctaac
      781 aaaaaagcca aaaacggcca gaatttagcg gacaatttac aagtctaaca ctgaaaatta
      841 catattgacc caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc
      901 gccaaccccg agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat
      961 aaaaccgacc aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc
     1021 tgtcgattca ggagagccta ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc
     1081 taccgcacac ctaccggctg gtggccccac cctgcattat aagccaacag aacgggtgac
     1141 gtcacgacac gacgagggcg cgcgctccca aaggtacggg tgcactgccc aacggcaccg
     1201 ccataactgc cgcccccgca acagacgaca aaccgagttc tccagtcagt gacaaacttc
     1261 acgtcagggt ccccagatgg tgcccagccc atctcacccg aataagagct ttcccgcatt
     1321 agcgaagggc tcaagacctt gggttcttgc cgcccaccat gccccccacc ttgtttcaac
     1381 gacctcacag cccgcctcac aagcgtcttc cattcaagac tcgggaacag ccgccatttt
     1441 gctgcgctcc ccccaacccc cagttcaggg caaccttgct cgcggaccca gactacagcc
     1501 cttggcggtc tctccacacg cttccgtccc accgagcggc ccggcggcca cgaaagcccc
     1561 ggccagccca gcagcccgct actcaccaag tgacgatcac agcgatccac aaacaagaac
     1621 cgcgacccaa atcccggctg cgacggaact agctgtgcca cacccggcgc gtccttatat
     1681 aatcatcggc gttcaccgcc ccacggagat ccctccgcag aatcgccgag aagggactac
     1741 ttttcctcgc ctgttccgct ctctggaaag aaaaccagtg ccctagagtc acccaagtcc
     1801 cgtcctaaaa tgtccttctg ctgatactgg ggttctaagg ccgagtctta tgagcagcgg
     1861 gccgctgtcc tgagcgtccg ggcggaagga tcaggacgct cctgcgccct tcgtctgacg
     1921 tggcagcgct cgccgtgacg aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag
     1981 gccaggcgcg ccccgatttc gatattcctc ggatctcgga tagatcggga ttgaggatgt
     2041 aggttgaact tctctcgggt gatcccgatc tcggatattc gggattggag taatacctgg
     2101 cgataggcgc tgccaagcaa tagccaaggc gcaaggccgg ccaagagact tgcgatagtt
     2161 cgttcgagag gtagcctgca agatggatcg gacgaaaggt ttgaagattt tccgacgagg
     2221 gccttggaga gttcggaaat ttaaatgcaa ctatggatga acgaaataga cagatcgctg
     2281 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2341 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2401 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2461 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2521 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2581 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2641 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2701 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2761 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2821 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2881 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2941 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3001 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3061 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3121 ttgctcacat gatttaaatg aaactcgtaa tcggagaacg ggtgcaatag caaggaccag
     3181 atcgaacttg actgacgcaa ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca
     3241 agtagctgac ttgactggtt aattaagttt cgatagccca aggtaaccaa tgagattagg
     3301 gagggaagaa agtttgatga actacttctg acccctagtg gtgtccagaa aagaccatta
     3361 aaggaatgtc ttgatcgaaa tcgagagatc gcctgcagga tagctgacga ccttaaggat
     3421 aaatttctgg taaggaggac acgtatggaa gtgggcaagt tggggaagcc gtatccgttg
     3481 ctgaatctgg catatgtggg agtataagac gcgcagcgtc gcatcaggca tttttttctg
     3541 cgccaatgcc ggtagatagc aatccggccc gaggggcaca aaaaacccct caagacccgt
     3601 ttagaggccc caaggggtta tgctacgata ccgatagaga tgggccaata ccacatttgt
     3661 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3721 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3781 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3841 caaactcatc aatgtatctt atcatgtctg tgagctgaag gtacgctgta tctcagtcag
     3901 tcaagctagc ttacgcaaac ccgtttaaaa acgattgtgc atcagccatg gtggcggccg
     3961 ctggcgggtg tgtcgagtgg atggtaggat cgacaaagat ctgatctgac ggttcactaa
     4021 accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     4081 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     4141 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     4201 attgatgtac tgccaaaacc gcatcagcat ggtaatagcg atgactaata cgtagatgta
     4261 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     4321 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     4381 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     4441 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     4501 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     4561 tgaccccgta attgattact agatctggct acactcgatc agcagttaga taataaaatc
     4621 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcaa cgattgatag
     4681 cataacccct tggggcctct aaacgggtct tgaggggttt tttgaattga gaggtagctg
     4741 aagctgctgt tatgaccact agagggaaga agatacctgt ggagctaatg gtccaagatg
     4801 tcgggagccg cgatcgctgc caaaaaggta agctgacgat agctgacgag attcgatgcc
     4861 cgacgagcaa atgagcgcca cggctgttcg cgtaagctcg aaacgacgga gattcgatag
     4921 agccagagtc gagtttaaac gattcgggat cttaaggata aatttctggt aaggaggaca
     4981 cgtatggaag tgggcaagtt ggggaagccg tatccgttgc tgaatctggc atatgtggga
     5041 gtataagacg cgcagcgtcg catcaggcat ttttttctgc gccaatgcac ctagaatcgg
     5101 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5161 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5221 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5281 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5341 gtctatcaac tgttgccggg aagctagagt aagtagttcg ccagttaata gtttacgcaa
     5401 ggttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5461 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5521 ggttagctcc ttcggtcctc caatcgttgt cagaagtaag ttggccgcag tgttatcact
     5581 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5641 tgtgactggt gaatactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5701 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5761 catcattgga aagcgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5821 cagttcgatg taacccactc gcgcacccaa ctgatcttca gcatctttta ctttcaccag
     5881 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5941 acggaaatgt tgaatactca tgcgaaagcc tcctgctgat cctgtctctt gatagatgtt
     6001 tgcaaaagct aggactcgat tatacgttga ccgatcagac tatcaatatc taggtttaaa
     6061 cagatcggta cgctgccgtg accaggcttg agtacaatat ctgacgattg gacttcctgg
     6121 gtcccaagct g
//
