LOCUS       sequence-297501-        6713 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 6713)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6713)
  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-m4252849
FEATURES             Location/Qualifiers
     source          1..6713
                     /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"
     promoter        2553..3762
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3599..3618
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3766..4365
                     /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(3766..3785)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4262..4282
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4392..4440
                     /note="synthetic polyadenylation signal"
     CDS             join(4706..4774,4775..5566)
                     /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(4924..4943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5579..5715
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5867..5938
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5943..5989)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6007..6055
                     /note="synthetic polyadenylation signal"
     enhancer        6136..6439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6441..6643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6594..6614
                     /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 ctggcctccg cgccgggttt tggcgcctcc
     2581 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     2641 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     2701 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     2761 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     2821 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     2881 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     2941 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     3001 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     3061 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     3121 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     3181 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     3241 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     3301 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     3361 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     3421 gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     3481 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     3541 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     3601 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     3661 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     3721 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     3781 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3841 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3901 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3961 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4021 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4081 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4141 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4201 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4261 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4321 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     4381 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     4441 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     4501 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     4561 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     4621 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     4681 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     4741 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4801 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     4861 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     4921 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4981 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     5041 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5101 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     5161 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5221 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     5281 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     5341 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5401 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5461 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5521 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     5581 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     5641 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     5701 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     5761 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     5821 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     5881 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     5941 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     6001 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     6061 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     6121 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     6181 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     6241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6421 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6481 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6541 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6601 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     6661 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
