LOCUS       sequence-311976-        4698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4698)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4698)
  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-m4133160
FEATURES             Location/Qualifiers
     source          1..4698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1003..1023
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1027..1092
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1145..1266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1156..1175)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1210..1229
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1381..1427
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1457..1593
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1638..1709
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1935..2523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2015..2034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3269..3337,3338..4129)
                     /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(3487..3506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4142..4278
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4430..4501
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4506..4552)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4570..4618
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggcagg gaatgcgaca
      601 gaggttcctg ccaactcaac ggtcttgtct ttttgtgctt tcgccgtgga cgctgcaaaa
      661 gcctacaagg attatctagc ctcgggtgga cagccaatca ctaactgcgt aaagatgctt
      721 tgtacccaca ccggcactgg ccaagcaatt acagtgaccc ccgaagctaa tatggaccag
      781 gagtccttcg gaggtgcgag ctgctgcctg tactgtcgtt gtcatataga tcacccgaat
      841 cccaaagggt tttgcgacct caagggcaaa tatgtccaga tcccaacgac atgcgccaac
      901 gatcctgtgg gattcactct gaagaacacc gtatgtaccg tttgcgggat gtggaagggg
      961 tatggctgta gttgcgacca attacgggaa ccgatgctgc aagaaaacct ttactttcag
     1021 tcgatggatt ataaggacca cgatggagac tacaaagatc atgacattga ttacaaggac
     1081 gacgatgaca aataagctag cttgactgac tgagatacag cgtaccttca gctcacagac
     1141 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1201 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1261 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1321 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
     1381 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
     1441 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     1501 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     1561 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
     1621 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
     1681 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
     1741 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     1801 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     1861 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2101 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2341 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     2761 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     2821 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     2881 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     2941 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     3001 atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac
     3061 gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc
     3121 aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag
     3181 atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca
     3241 agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat
     3301 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3361 aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag
     3421 cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa
     3481 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3541 ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct
     3601 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3661 tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca
     3721 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3781 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact
     3841 attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc
     3901 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3961 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4021 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4081 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg
     4141 tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     4201 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     4261 ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct
     4321 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
     4381 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
     4441 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
     4501 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
     4561 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
     4621 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
     4681 cccatatatg gagttccg
//
