LOCUS       sequence-304504-        4323 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 4323)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4323)
  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-m4133342
FEATURES             Location/Qualifiers
     source          1..4323
                     /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"
     CDS             join(2316..2384,2385..3176)
                     /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(2534..2553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3189..3325
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3477..3548
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3553..3599)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3617..3665
                     /note="synthetic polyadenylation signal"
     enhancer        3746..4049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4051..4253
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4204..4224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4322..4323,1..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 gtgtaccgag aacgatcctc tcagtgcgag
     2101 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     2161 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     2221 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     2281 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     2341 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2401 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     2461 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     2521 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2581 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     2641 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2701 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     2761 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2821 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     2881 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     2941 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3001 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3061 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3121 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     3181 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     3241 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     3301 ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat
     3361 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     3421 tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc
     3481 ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc
     3541 tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat
     3601 caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     3661 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
     3721 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3781 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3841 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3901 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     3961 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4021 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
     4081 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4141 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4201 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     4261 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     4321 cgc
//
