LOCUS       sequence-295359-        4674 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 4674)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4674)
  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-m4134110
FEATURES             Location/Qualifiers
     source          1..4674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..445
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             446..490
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    543..664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(554..573)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     608..627
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      779..825
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      855..991
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1036..1107
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1333..1921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1413..1432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2667..2735,2736..3527)
                     /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(2885..2904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3540..3676
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3828..3899
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3904..3950)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3968..4016
                     /note="synthetic polyadenylation signal"
     enhancer        4097..4400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4402..4604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4555..4575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4673..4674,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gggaatgcga cagaggttcc tgccaactca acggtcttgt ctttttgtgc
       61 tttcgccgtg gacgctgcaa aagcctacaa ggattatcta gcctcgggtg gacagccaat
      121 cactaactgc gtaaagatgc tttgtaccca caccggcact ggccaagcaa ttacagtgac
      181 ccccgaagct aatatggacc aggagtcctt cggaggtgcg agctgctgcc tgtactgtcg
      241 ttgtcatata gatcacccga atcccaaagg gttttgcgac ctcaagggca aatatgtcca
      301 gatcccaacg acatgcgcca acgatcctgt gggattcact ctgaagaaca ccgtatgtac
      361 cgtttgcggg atgtggaagg ggtatggctg tagttgcgac caattacggg aaccgatgct
      421 gcaagaaaac ctttactttc agtcgaaaga aaccgcggca gctaagttcg agaggcagca
      481 catggacagt taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
      541 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      601 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      661 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
      721 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
      781 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
      841 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
      901 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
      961 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     1021 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     1081 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     1141 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     1201 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1261 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1741 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2041 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2101 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     2161 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     2221 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2281 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2341 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2401 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     2461 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     2521 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     2581 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     2641 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     2701 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2761 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     2821 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     2881 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2941 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     3001 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3061 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     3121 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3181 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     3241 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     3301 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3361 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3421 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3481 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     3541 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     3601 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     3661 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     3721 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     3781 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     3841 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     3901 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     3961 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     4021 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     4081 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4141 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4201 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4261 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4321 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4381 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     4441 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4501 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4561 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     4621 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
