LOCUS       sequence-300221-        4806 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 4806)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4806)
  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-m4133457
FEATURES             Location/Qualifiers
     source          1..4806
                     /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             101..178
                     /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"
     CDS             206..226
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    675..796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(686..705)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     740..759
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      911..957
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      987..1123
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1168..1239
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1465..2053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1545..1564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2799..2867,2868..3659)
                     /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(3017..3036)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3672..3808
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3960..4031
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4036..4082)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4100..4148
                     /note="synthetic polyadenylation signal"
     enhancer        4229..4532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4534..4736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4687..4707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcggga cggcgaatcc cggggctgat aaatccctgg aaaaggagat ggaagaagaa
      121 ctttattgca gtgtctgcgg ctaaccggtt caaaaagatc agttcaagcg gcgccctaga
      181 ttacgacatt cctacgaccg cctccgagaa tctctatttc cagggtgaat ttggcttggc
      241 acaacatgac gaggccgtcg ataacaagtt taataaagaa cagcagaacg ctttctatga
      301 gatccttcac ttaccaaatc tgaacgaaga gcaacgcaac gctttcattc agagcctgaa
      361 agacgatccc tcgcagtctg caaatctcct ggccgaagcc aagaagctga atgacgcaca
      421 ggctccaaaa gttgataaca agtttaacaa ggagcagcag aatgctttct acgagatatt
      481 gcaccttcct aatctgaatg aggaacaaag aaatgcattt atccagtccc taaaagacga
      541 tccatcacag agtgccaacc tgcttgcgga agccaaaaag ctcaatgatg cccaggcccc
      601 gaaagtagac gcgaaccatc aataagctag cttgactgac tgagatacag cgtaccttca
      661 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      721 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      781 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
      841 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
      901 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
      961 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     1021 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     1081 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     1141 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     1201 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     1261 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     1321 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1381 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1441 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1501 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1561 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1621 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1681 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1741 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1801 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1861 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1921 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1981 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2041 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2101 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2161 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2221 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2281 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     2341 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2401 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2461 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     2521 gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     2581 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     2641 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     2701 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     2761 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     2821 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2881 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     2941 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     3001 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3061 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     3121 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3181 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     3241 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3301 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     3361 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     3421 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3481 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3541 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3601 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3661 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     3721 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     3781 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc
     3841 tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg
     3901 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac
     3961 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc
     4021 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     4081 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg
     4141 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat
     4201 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4261 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4321 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4381 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4441 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4501 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg
     4561 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     4621 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     4681 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     4741 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc
     4801 ggccgc
//
