LOCUS       sequence-307255-        6599 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 6599)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6599)
  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-m4134156
FEATURES             Location/Qualifiers
     source          1..6599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             419..439
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             443..508
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    561..682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(572..591)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     626..645
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      797..843
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      873..1009
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1054..1125
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1351..1939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1431..1450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2439..3648
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3485..3504
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3652..4251
                     /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(3652..3671)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4148..4168
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4278..4326
                     /note="synthetic polyadenylation signal"
     CDS             join(4592..4660,4661..5452)
                     /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(4810..4829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5465..5601
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5753..5824
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5829..5875)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5893..5941
                     /note="synthetic polyadenylation signal"
     enhancer        6022..6325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6327..6529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6480..6500
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6598..6599,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggt aatgcaacag aagtgcctgc caattcaact gtattatctt tctgtgcttt
       61 tgctgtagat gctgctaaag cttacaaaga ttatctagct agtgggggac aaccaatcac
      121 taattgtgtt aagatgttgt gtacacacac tggtactggt caggcaataa cagttacacc
      181 ggaagccaat atggatcaag aatcctttgg tggtgcatcg tgttgtctgt actgccgttg
      241 ccacatagat catccaaatc ctaaaggatt ttgtgactta aaaggtaagt atgtacaaat
      301 acctacaact tgtgctaatg accctgtggg ttttacactt aaaaacacag tctgtaccgt
      361 ctgcggtatg tggaaaggtt atggctgtag ttgtgatcaa ctccgcgaac ccatgcttga
      421 aaacctttac tttcagtcga tggattataa ggaccacgat ggagactaca aagatcatga
      481 cattgattac aaggacgacg atgacaaata agctagcttg actgactgag atacagcgta
      541 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      601 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      661 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      721 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
      781 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      841 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
      901 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
      961 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     1021 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     1081 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     1141 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     1201 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     1261 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1321 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1381 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1441 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1501 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1561 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1621 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1681 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1741 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1801 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1861 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1921 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1981 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2041 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2101 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2161 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     2221 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     2281 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2341 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2401 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     2461 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     2521 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     2581 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     2641 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     2701 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     2761 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     2821 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     2881 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     2941 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     3001 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     3061 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     3121 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     3181 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     3241 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     3301 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     3361 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     3421 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     3481 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     3541 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     3601 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag
     3661 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3721 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3781 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3841 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3901 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3961 gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4021 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4081 gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4141 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4201 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg
     4261 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     4321 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     4381 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     4441 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     4501 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     4561 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     4621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4681 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     4741 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     4801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4861 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     4921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4981 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     5041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     5161 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     5221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5281 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     5461 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     5521 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     5581 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     5641 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     5701 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     5761 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     5821 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     5881 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     5941 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     6001 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6061 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6121 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6181 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6241 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6301 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     6361 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6421 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6481 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     6541 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
