LOCUS       sequence-300695-        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"
     terminator      complement(75..121)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    139..187
                     /note="synthetic polyadenylation signal"
     enhancer        268..571
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        573..775
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     726..746
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             892..912
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             913..990
                     /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    1043..1164
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1054..1073)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1108..1127
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1279..1325
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1355..1491
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1536..1607
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1833..2421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1913..1932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3167..3235,3236..4027)
                     /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(3385..3404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4040..4176
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(4328..4329,1..70)
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
       61 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
      121 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
      181 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
      241 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      301 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      361 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      421 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      481 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      541 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
      601 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      661 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      721 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      781 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
      841 gccgccacca tgagtgccga tgctcagtcc tttctcaacg gtttcgcagt ggaaaacctt
      901 tactttcagt cgaagaggcg atggaaaaag aattttatcg cggtgagtgc agccaacaga
      961 ttcaaaaaga tatcctcgtc tggtgccctc taagctagct tgactgactg agatacagcg
     1021 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1081 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1141 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1201 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     1261 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1321 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     1381 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     1441 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     1501 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     1561 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     1621 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     1681 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     1741 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     1801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2461 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2521 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2581 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2641 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     2701 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     2761 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     2821 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     2881 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct
     2941 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     3001 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     3061 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     3121 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     3181 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3241 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     3301 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     3361 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3421 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     3481 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3541 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     3601 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3661 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3721 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     3781 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3841 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3901 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3961 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4021 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     4081 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     4141 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     4201 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     4261 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     4321 ctcccgaca
//
