LOCUS       sequence-300571-        6236 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 6236)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6236)
  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-m4252799
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    92..140
                     /note="synthetic polyadenylation signal"
     terminator      158..204
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    209..280
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(432..568)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(581..1372,1373..1441))
                     /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     1204..1223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1707..1755
                     /note="synthetic polyadenylation signal"
     CDS             complement(1782..2381)
                     /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(1865..1885)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2362..2381
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2385..3594)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2529..2548)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4094..4682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4583..4602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4908..4979
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5024..5160)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5190..5236)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5351..5472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5388..5407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5442..5461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5525..5602)
                     /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(5603..5623)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5740..5942)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5769..5789)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(5944..6236,1..11)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
       61 agatctggct acactcgatc agcagttaga taataaaatc gctatccatc gaagatggat
      121 gtgtgttggt tttttgtgtg tgtaacgcaa cgattgatag cataacccct tggggcctct
      181 aaacgggtct tgaggggttt tttgaattga gaggtagctg aagctgctgt tatgaccact
      241 agagggaaga agatacctgt ggagctaatg gtccaagatg tcgggagccg cgatcgctgc
      301 caaaaaggta agctgacgat agctgacgag attcgatgcc cgacgagcaa atgagcgcca
      361 cggctgttcg cgtaagctcg aaacgacgga gattcgatag agccagagtc gagtttaaac
      421 gattcgggat cttaaggata aatttctggt aaggaggaca cgtatggaag tgggcaagtt
      481 ggggaagccg tatccgttgc tgaatctggc atatgtggga gtataagacg cgcagcgtcg
      541 catcaggcat ttttttctgc gccaatgcac ctagaatcgg ttaccaatgc ttaatcagtg
      601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
      781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctatcaac tgttgccggg
      841 aagctagagt aagtagttcg ccagttaata gtttacgcaa ggttgttgcc attgctacag
      901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
      961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1021 caatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gaatactcaa
     1141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aagcgttctt
     1261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1321 gcgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1441 tgcgaaagcc tcctgctgat cctgtctctt gatagatgtt tgcaaaagct aggactcgat
     1501 tatacgttga ccgatcagac tatcaatatc taggtttaaa cagatcggta cgctgccgtg
     1561 accaggcttg agtacaatat ctgacgattg gacttcctgg gtcccaagct ggattccgac
     1621 tggctgactg ccaagcaacg atatggatcg tcgagactcg cactgagagg atcgttctcg
     1681 gtacaccagg cgcgcctagc gttacacaca caaaaaacca acacacatcc atcttcgatg
     1741 gatagcgatt ttattatcta agactaggct tacatactag cctaggcacc gggcttgcgg
     1801 gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg
     1861 agccgctcgt agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg
     1921 gcgcgctcgg cggcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg
     1981 tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc
     2041 gccaggaggc cctccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg
     2101 cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc
     2161 gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg
     2221 aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg
     2281 cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg
     2341 tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca tgttgtctaa caaaaaagcc
     2401 aaaaacggcc agaatttagc ggacaattta caagtctaac actgaaaatt acatattgac
     2461 ccaaatgatt acatttcaaa aggtgcctaa aaaacttcac aaaacacact cgccaacccc
     2521 gagcgcatag ttcaaaaccg gagcttcagc tacttaagaa gataggtaca taaaaccgac
     2581 caaagaaact gacgcctcac ttatccctcc cctcaccaga ggtccggcgc ctgtcgattc
     2641 aggagagcct accctaggcc cgaaccctgc gtcctgcgac ggagaaaagc ctaccgcaca
     2701 cctaccggct ggtggcccca ccctgcatta taagccaaca gaacgggtga cgtcacgaca
     2761 cgacgagggc gcgcgctccc aaaggtacgg gtgcactgcc caacggcacc gccataactg
     2821 ccgcccccgc aacagacgac aaaccgagtt ctccagtcag tgacaaactt cacgtcaggg
     2881 tccccagatg gtgcccagcc catctcaccc gaataagagc tttcccgcat tagcgaaggg
     2941 ctcaagacct tgggttcttg ccgcccacca tgccccccac cttgtttcaa cgacctcaca
     3001 gcccgcctca caagcgtctt ccattcaaga ctcgggaaca gccgccattt tgctgcgctc
     3061 cccccaaccc ccagttcagg gcaaccttgc tcgcggaccc agactacagc ccttggcggt
     3121 ctctccacac gcttccgtcc caccgagcgg cccggcggcc acgaaagccc cggccagccc
     3181 agcagcccgc tactcaccaa gtgacgatca cagcgatcca caaacaagaa ccgcgaccca
     3241 aatcccggct gcgacggaac tagctgtgcc acacccggcg cgtccttata taatcatcgg
     3301 cgttcaccgc cccacggaga tccctccgca gaatcgccga gaagggacta cttttcctcg
     3361 cctgttccgc tctctggaaa gaaaaccagt gccctagagt cacccaagtc ccgtcctaaa
     3421 atgtccttct gctgatactg gggttctaag gccgagtctt atgagcagcg ggccgctgtc
     3481 ctgagcgtcc gggcggaagg atcaggacgc tcctgcgccc ttcgtctgac gtggcagcgc
     3541 tcgccgtgac gaggggggcg cccgcgggag gcgccaaaac ccggcgcgga ggccaggcgc
     3601 gccccgattt cgatattcct cggatctcgg atagatcggg attgaggatg taggttgaac
     3661 ttctctcggg tgatcccgat ctcggatatt cgggattgga gtaatacctg gcgataggcg
     3721 ctgccaagca atagccaagg cgcaaggccg gccaagagac ttgcgatagt tcgttcgaga
     3781 ggtagcctgc aagatggatc ggacgaaagg tttgaagatt ttccgacgag ggccttggag
     3841 agttcggaaa tttaaatgca actatggatg aacgaaatag acagatcgct gagataggtg
     3901 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3961 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4021 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4081 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4141 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4201 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4261 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4321 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4381 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4441 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4501 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4561 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4621 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4681 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4741 tgatttaaat gaaactcgta atcggagaac gggtgcaata gcaaggacca gatcgaactt
     4801 gactgacgca attctgagcg tagtcgaaac ccagacaagt cgatctcgcc aagtagctga
     4861 cttgactggt taattaagtt tcgatagccc aaggtaacca atgagattag ggagggaaga
     4921 aagtttgatg aactacttct gacccctagt ggtgtccaga aaagaccatt aaaggaatgt
     4981 cttgatcgaa atcgagagat cgcctgcagg atagctgacg accttaagga taaatttctg
     5041 gtaaggagga cacgtatgga agtgggcaag ttggggaagc cgtatccgtt gctgaatctg
     5101 gcatatgtgg gagtataaga cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc
     5161 cggtagatag caatccggcc cgaggggcac aaaaaacccc tcaagacccg tttagaggcc
     5221 ccaaggggtt atgctacgat accgatagag atgggccaat accacatttg tagaggtttt
     5281 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     5341 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5401 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5461 caatgtatct tatcatgtct gtgagctgaa ggtacgctgt atctcagtca gtcaagctag
     5521 cttagagggc accagacgag gatatctttt tgaatctgtt ggctgcactc accgcgataa
     5581 aattcttttt ccatcgcctc ttcgactgaa agtaaaggtt ttccactgcg aaaccgttga
     5641 gaaaggactg agcatcggca ctcatggtgg cggccgctgg cgggtgtgtc gagtggatgg
     5701 taggatcgac aaagatctga tctgacggtt cactaaacca gctctgctta tatagacctc
     5761 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     5821 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     5881 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     5941 cagcatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     6001 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     6061 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     6121 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     6181 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaa
//
