LOCUS       sequence-297379-        4329 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 4329)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4329)
  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-m4133333
FEATURES             Location/Qualifiers
     source          1..4329
                     /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..145
                     /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    198..319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(209..228)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     263..282
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      434..480
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      510..646
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    691..762
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(988..1576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1068..1087)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2322..2390,2391..3182)
                     /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(2540..2559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3195..3331
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3483..3554
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3559..3605)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3623..3671
                     /note="synthetic polyadenylation signal"
     enhancer        3752..4055
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4057..4259
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4210..4230
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgaag aggcgatgga aaaagaattt tatcgcggtg agtgcagcca acagattcaa
      121 aaagatatcc tcgtctggtg ccctctaagc tagcttgact gactgagata cagcgtacct
      181 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      241 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      301 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
      361 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
      421 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      481 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
      541 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
      601 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
      661 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
      721 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
      781 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
      841 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
      901 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
      961 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1021 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1081 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1141 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1201 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1261 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1321 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1381 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1441 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1501 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1561 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1621 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1681 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1741 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1801 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     1861 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     1921 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     1981 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     2041 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag
     2101 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     2161 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     2221 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     2281 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     2341 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2401 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     2461 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     2521 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2581 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     2641 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2701 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     2761 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2821 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2881 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     2941 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3001 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3061 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3181 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     3241 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     3301 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     3361 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     3421 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     3481 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     3541 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     3601 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     3661 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     3721 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     3781 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     3841 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     3901 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     3961 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4021 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     4081 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4141 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4201 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     4261 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     4321 agcggccgc
//
