LOCUS       sequence-307340-        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"
     misc_feature    2..73
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(225..361)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(374..1165,1166..1234))
                     /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     997..1016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1500..1548
                     /note="synthetic polyadenylation signal"
     CDS             complement(1575..2174)
                     /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(1658..1678)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2155..2174
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2178..3387)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2322..2341)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      3887..4475
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4376..4395
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4701..4772
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4817..4953)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(4983..5029)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5144..5265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5181..5200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5235..5254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5714..5734)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(5762..5839)
                     /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             complement(5873..5893)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6010..6212)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6039..6059)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6214..6517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    6598..6646
                     /note="synthetic polyadenylation signal"
     terminator      6664..6710
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tgagaggtag ctgaagctgc tgttatgacc actagaggga agaagatacc tgtggagcta
       61 atggtccaag atgtcgggag ccgcgatcgc tgccaaaaag gtaagctgac gatagctgac
      121 gagattcgat gcccgacgag caaatgagcg ccacggctgt tcgcgtaagc tcgaaacgac
      181 ggagattcga tagagccaga gtcgagttta aacgattcgg gatcttaagg ataaatttct
      241 ggtaaggagg acacgtatgg aagtgggcaa gttggggaag ccgtatccgt tgctgaatct
      301 ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg catttttttc tgcgccaatg
      361 cacctagaat cggttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      421 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      481 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
      541 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
      601 ccgcctccat ccagtctatc aactgttgcc gggaagctag agtaagtagt tcgccagtta
      661 atagtttacg caaggttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
      721 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
      781 tgtgcaaaaa agcggttagc tccttcggtc ctccaatcgt tgtcagaagt aagttggccg
      841 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
      901 taagatgctt ttctgtgact ggtgaatact caaccaagtc attctgagaa tagtgtatgc
      961 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1021 ctttaaaagt gctcatcatt ggaaagcgtt cttcggggcg aaaactctca aggatcttac
     1081 cgctgttgag atccagttcg atgtaaccca ctcgcgcacc caactgatct tcagcatctt
     1141 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1201 gaataagggc gacacggaaa tgttgaatac tcatgcgaaa gcctcctgct gatcctgtct
     1261 cttgatagat gtttgcaaaa gctaggactc gattatacgt tgaccgatca gactatcaat
     1321 atctaggttt aaacagatcg gtacgctgcc gtgaccaggc ttgagtacaa tatctgacga
     1381 ttggacttcc tgggtcccaa gctggattcc gactggctga ctgccaagca acgatatgga
     1441 tcgtcgagac tcgcactgag aggatcgttc tcggtacacc aggcgcgcct agcgttacac
     1501 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaagactag
     1561 gcttacatac tagcctaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     1621 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     1681 ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggcggcctc cactccgggg
     1741 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     1801 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccctccat ctgttgctgc
     1861 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     1921 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     1981 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     2041 cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg
     2101 gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc
     2161 ttgtactcgg tcatgttgtc taacaaaaaa gccaaaaacg gccagaattt agcggacaat
     2221 ttacaagtct aacactgaaa attacatatt gacccaaatg attacatttc aaaaggtgcc
     2281 taaaaaactt cacaaaacac actcgccaac cccgagcgca tagttcaaaa ccggagcttc
     2341 agctacttaa gaagataggt acataaaacc gaccaaagaa actgacgcct cacttatccc
     2401 tcccctcacc agaggtccgg cgcctgtcga ttcaggagag cctaccctag gcccgaaccc
     2461 tgcgtcctgc gacggagaaa agcctaccgc acacctaccg gctggtggcc ccaccctgca
     2521 ttataagcca acagaacggg tgacgtcacg acacgacgag ggcgcgcgct cccaaaggta
     2581 cgggtgcact gcccaacggc accgccataa ctgccgcccc cgcaacagac gacaaaccga
     2641 gttctccagt cagtgacaaa cttcacgtca gggtccccag atggtgccca gcccatctca
     2701 cccgaataag agctttcccg cattagcgaa gggctcaaga ccttgggttc ttgccgccca
     2761 ccatgccccc caccttgttt caacgacctc acagcccgcc tcacaagcgt cttccattca
     2821 agactcggga acagccgcca ttttgctgcg ctccccccaa cccccagttc agggcaacct
     2881 tgctcgcgga cccagactac agcccttggc ggtctctcca cacgcttccg tcccaccgag
     2941 cggcccggcg gccacgaaag ccccggccag cccagcagcc cgctactcac caagtgacga
     3001 tcacagcgat ccacaaacaa gaaccgcgac ccaaatcccg gctgcgacgg aactagctgt
     3061 gccacacccg gcgcgtcctt atataatcat cggcgttcac cgccccacgg agatccctcc
     3121 gcagaatcgc cgagaaggga ctacttttcc tcgcctgttc cgctctctgg aaagaaaacc
     3181 agtgccctag agtcacccaa gtcccgtcct aaaatgtcct tctgctgata ctggggttct
     3241 aaggccgagt cttatgagca gcgggccgct gtcctgagcg tccgggcgga aggatcagga
     3301 cgctcctgcg cccttcgtct gacgtggcag cgctcgccgt gacgaggggg gcgcccgcgg
     3361 gaggcgccaa aacccggcgc ggaggccagg cgcgccccga tttcgatatt cctcggatct
     3421 cggatagatc gggattgagg atgtaggttg aacttctctc gggtgatccc gatctcggat
     3481 attcgggatt ggagtaatac ctggcgatag gcgctgccaa gcaatagcca aggcgcaagg
     3541 ccggccaaga gacttgcgat agttcgttcg agaggtagcc tgcaagatgg atcggacgaa
     3601 aggtttgaag attttccgac gagggccttg gagagttcgg aaatttaaat gcaactatgg
     3661 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3721 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3781 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3841 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3901 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3961 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4021 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4081 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4141 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4201 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4261 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4321 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4381 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4441 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4501 ggttcctggc cttttgctgg ccttttgctc acatgattta aatgaaactc gtaatcggag
     4561 aacgggtgca atagcaagga ccagatcgaa cttgactgac gcaattctga gcgtagtcga
     4621 aacccagaca agtcgatctc gccaagtagc tgacttgact ggttaattaa gtttcgatag
     4681 cccaaggtaa ccaatgagat tagggaggga agaaagtttg atgaactact tctgacccct
     4741 agtggtgtcc agaaaagacc attaaaggaa tgtcttgatc gaaatcgaga gatcgcctgc
     4801 aggatagctg acgaccttaa ggataaattt ctggtaagga ggacacgtat ggaagtgggc
     4861 aagttgggga agccgtatcc gttgctgaat ctggcatatg tgggagtata agacgcgcag
     4921 cgtcgcatca ggcatttttt tctgcgccaa tgccggtaga tagcaatccg gcccgagggg
     4981 cacaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctac gataccgata
     5041 gagatgggcc aataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     5101 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     5161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtgagct
     5281 gaaggtacgc tgtatctcag tcagtcaagc tagcttattg atggttcgcg tctactttcg
     5341 gggcctgggc atcattgagc tttttggctt ccgcaagcag gttggcactc tgtgatggat
     5401 cgtcttttag ggactggata aatgcatttc tttgttcctc attcagatta ggaaggtgca
     5461 atatctcgta gaaagcattc tgctgctcct tgttaaactt gttatcaact tttggagcct
     5521 gtgcgtcatt cagcttcttg gcttcggcca ggagatttgc agactgcgag ggatcgtctt
     5581 tcaggctctg aatgaaagcg ttgcgttgct cttcgttcag atttggtaag tgaaggatct
     5641 catagaaagc gttctgctgt tctttattaa acttgttatc gacggcctcg tcatgttgtg
     5701 ccaagccaaa ttcaccctgg aaatagagat tctcggaggc ggtcgtagga atgtcgtaat
     5761 ctagggcgcc gcttgaactg atctttttga accggttagc cgcagacact gcaataaagt
     5821 tcttcttcca tctccttttc cagggattta tcagccccgg gattcgccgt cccgactgaa
     5881 agtaaaggtt ttccactgcg aaaccgttga gaaaggactg agcatcggca ctcatggtgg
     5941 cggccgctgg cgggtgtgtc gagtggatgg taggatcgac aaagatctga tctgacggtt
     6001 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     6061 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     6121 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     6181 acgcccattg atgtactgcc aaaaccgcat cagcatggta atagcgatga ctaatacgta
     6241 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     6301 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     6361 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     6421 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     6481 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     6541 aactaatgac cccgtaattg attactagat ctggctacac tcgatcagca gttagataat
     6601 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgcaacgat
     6661 tgatagcata accccttggg gcctctaaac gggtcttgag gggttttttg aat
//
