LOCUS       sequence-297473-        6230 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 6230)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6230)
  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-m4252783
FEATURES             Location/Qualifiers
     source          1..6230
                     /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..139
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    192..313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(203..222)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     257..276
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      428..474
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      504..640
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    685..756
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(982..1570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1062..1081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2070..3279
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3116..3135
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3283..3882
                     /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(3283..3302)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3779..3799
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3909..3957
                     /note="synthetic polyadenylation signal"
     CDS             join(4223..4291,4292..5083)
                     /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(4441..4460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5096..5232
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5384..5455
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5460..5506)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5524..5572
                     /note="synthetic polyadenylation signal"
     enhancer        5653..5956
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5958..6160
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6111..6131
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6229..6230,1..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 gaagaggcga tggaaaaaga attttatcgc ggtgagtgca gccaacagat tcaaaaagat
      121 atcctcgtct ggtgccctct aagctagctt gactgactga gatacagcgt accttcagct
      181 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      241 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      301 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
      361 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
      421 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
      481 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
      541 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
      601 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
      661 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
      721 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
      781 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
      841 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
      901 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1801 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     1861 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     1921 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     1981 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     2041 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     2101 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     2161 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     2221 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     2281 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     2341 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     2401 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     2461 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     2521 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     2581 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     2641 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     2701 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     2761 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     2821 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     2881 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     2941 gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     3001 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     3061 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     3121 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     3181 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     3241 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc
     3301 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3361 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3421 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3481 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3541 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3601 caacagatgg agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3661 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3721 ggagtggagg ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3781 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3841 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag
     3901 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     3961 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     4021 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     4081 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     4141 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     4201 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4261 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4321 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     4381 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     4441 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4501 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     4561 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4621 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     4681 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4741 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     4801 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     4861 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4921 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4981 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5041 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     5101 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     5161 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     5221 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     5281 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     5341 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     5401 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     5461 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     5521 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     5581 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     5641 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5701 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5761 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5821 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5881 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5941 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     6001 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     6061 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     6121 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     6181 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
