LOCUS       sequence-295205-        6700 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 6700)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6700)
  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-m4133621
FEATURES             Location/Qualifiers
     source          1..6700
                     /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"
     promoter        2547..3756
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3593..3612
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3760..4359
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3760..3779)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4256..4276
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4386..4434
                     /note="synthetic polyadenylation signal"
     CDS             join(4700..4768,4769..5560)
                     /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(4918..4937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5573..5709
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5854..5925
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5930..5976)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5994..6042
                     /note="synthetic polyadenylation signal"
     enhancer        6123..6426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6428..6630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6581..6601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6699..6700,1..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 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     2581 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     2641 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     2701 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     2761 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     2821 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     2881 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     2941 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     3001 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     3061 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     3121 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     3181 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     3241 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     3301 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     3361 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     3421 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     3481 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     3541 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     3601 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     3661 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     3721 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     3781 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3841 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3901 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3961 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4021 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4081 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4141 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4201 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     4261 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4321 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     4381 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     4441 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     4501 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     4561 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     4621 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     4681 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     4741 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4801 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4861 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     4921 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4981 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5041 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5101 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5161 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5221 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     5281 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     5341 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5401 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5461 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5521 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     5581 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     5641 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     5701 tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     5761 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     5821 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     5881 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     5941 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     6001 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     6061 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6421 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     6481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     6541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     6601 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
     6661 tcgatcctac catccactcg acacacccgc cagcggccgc
//
