LOCUS       sequence-379430-        6554 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 6554)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6554)
  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-m4134309
FEATURES             Location/Qualifiers
     source          1..6554
                     /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..463
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    516..637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(527..546)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     581..600
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      752..798
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      828..964
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1009..1080
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1306..1894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1386..1405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2394..3603
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3440..3459
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3607..4206
                     /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(3607..3626)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4103..4123
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4233..4281
                     /note="synthetic polyadenylation signal"
     CDS             join(4547..4615,4616..5407)
                     /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(4765..4784)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5420..5556
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5708..5779
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5784..5830)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5848..5896
                     /note="synthetic polyadenylation signal"
     enhancer        5977..6280
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6282..6484
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6435..6455
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6553..6554,1..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 agtcgcacca tcaccatcac cattaagcta gcttgactga
      481 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
      541 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      601 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      661 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      721 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
      781 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
      841 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
      901 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
      961 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     1021 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     1081 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     1141 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     1201 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     1261 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1321 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1381 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1441 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1501 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1561 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1621 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1681 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1741 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1801 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1861 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1921 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2101 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     2161 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     2221 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     2281 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     2341 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     2401 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     2461 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     2521 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     2581 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     2641 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     2701 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     2761 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     2821 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     2881 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     2941 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     3001 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     3061 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     3121 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     3181 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     3241 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     3301 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     3361 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     3421 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     3481 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     3541 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     3601 gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3661 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     3721 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     3781 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     3841 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     3901 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
     3961 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4021 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
     4081 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4141 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4201 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     4261 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     4321 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     4381 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     4441 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     4501 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     4561 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4621 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     4681 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     4741 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4801 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     4861 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4921 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     4981 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5041 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5101 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     5161 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5221 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5281 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5341 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5401 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     5461 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     5521 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     5581 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     5641 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     5701 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     5761 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     5821 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     5881 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     5941 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6001 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6061 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6121 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6181 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6241 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     6301 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     6361 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     6421 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     6481 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     6541 ccgccagcgg ccgc
//
