LOCUS       sequence-295917-        4800 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 4800)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4800)
  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-m4133495
FEATURES             Location/Qualifiers
     source          1..4800
                     /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             95..172
                     /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             200..220
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    669..790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(680..699)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     734..753
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      905..951
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      981..1117
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1162..1233
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1459..2047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1539..1558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2793..2861,2862..3653)
                     /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(3011..3030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3666..3802
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3954..4025
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4030..4076)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4094..4142
                     /note="synthetic polyadenylation signal"
     enhancer        4223..4526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4528..4730
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4681..4701
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4799..4800,1..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 gggacggcga atcccggggc tgataaatcc ctggaaaagg agatggaaga agaactttat
      121 tgcagtgtct gcggctaacc ggttcaaaaa gatcagttca agcggcgccc tagattacga
      181 cattcctacg accgcctccg agaatctcta tttccagggt gaatttggct tggcacaaca
      241 tgacgaggcc gtcgataaca agtttaataa agaacagcag aacgctttct atgagatcct
      301 tcacttacca aatctgaacg aagagcaacg caacgctttc attcagagcc tgaaagacga
      361 tccctcgcag tctgcaaatc tcctggccga agccaagaag ctgaatgacg cacaggctcc
      421 aaaagttgat aacaagttta acaaggagca gcagaatgct ttctacgaga tattgcacct
      481 tcctaatctg aatgaggaac aaagaaatgc atttatccag tccctaaaag acgatccatc
      541 acagagtgcc aacctgcttg cggaagccaa aaagctcaat gatgcccagg ccccgaaagt
      601 agacgcgaac catcaataag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      661 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      721 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      781 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      841 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
      901 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
      961 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1021 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1081 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1141 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     1201 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     1261 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     1321 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     1381 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1441 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1501 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1561 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1621 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1681 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1741 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1801 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1861 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1921 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1981 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2041 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2101 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2161 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2221 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2281 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     2341 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     2401 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2461 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2521 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     2581 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     2641 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     2701 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     2761 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     2821 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2881 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     2941 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     3001 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3061 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     3121 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3181 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     3241 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3301 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     3361 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     3421 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3481 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3541 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3601 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     3661 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     3721 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     3781 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     3841 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     3901 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     3961 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     4021 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     4081 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     4141 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     4201 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4261 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4321 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4381 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4441 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4501 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     4561 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4621 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4681 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     4741 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
