LOCUS       sequence-295598-        6919 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 6919)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6919)
  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-m4134092
FEATURES             Location/Qualifiers
     source          1..6919
                     /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             794..811
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    888..1009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(899..918)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     953..972
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1124..1170
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1200..1336
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1381..1452
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1678..2266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1758..1777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2766..3975
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3812..3831
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3979..4578
                     /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(3979..3998)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4475..4495
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4605..4653
                     /note="synthetic polyadenylation signal"
     CDS             join(4919..4987,4988..5779)
                     /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(5137..5156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5792..5928
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6073..6144
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6149..6195)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6213..6261
                     /note="synthetic polyadenylation signal"
     enhancer        6342..6645
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6647..6849
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6800..6820
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6918..6919,1..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 agtcgatgag tgacaaaatc attcatctaa ccgatgactc
      481 ctttgataca gacgtgttga aggccgatgg tgcgatactg gtagatttct gggctgaatg
      541 gtgtggcccg tgcaagatga tcgcacccat tctggacgag atcgccgacg agtatcaagg
      601 aaaattaact gtcgctaagc ttaatataga tcagaaccca gggacggcac ctaaatacgg
      661 catcagggga atacctaccc tcctgctctt taagaatggg gaagttgccg cgacaaaagt
      721 gggagccctg tctaagggcc agctaaaaga gttcttggac gctaacctgg caggttcagg
      781 ctcggggcac atgcaccatc accatcacca tagcagcggg ctcgtgccac gcggctaagc
      841 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      901 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      961 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1021 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1081 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1141 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1201 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1261 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1321 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     1381 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1441 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1501 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1561 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2101 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2521 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2581 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2641 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2701 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2761 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     2821 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     2881 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     2941 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     3001 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     3061 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     3121 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     3181 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     3241 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     3301 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     3361 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     3421 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     3481 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     3541 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     3601 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
     3661 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     3721 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     3781 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     3841 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     3901 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     3961 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4021 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4081 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4141 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4201 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4261 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
     4321 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     4381 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
     4441 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4501 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4561 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     4621 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     4681 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     4741 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     4801 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     4861 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     4921 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4981 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     5041 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5101 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5161 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5221 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5281 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     5341 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5401 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5461 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     5521 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5581 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5641 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5701 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5761 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     5821 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5881 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcgtttaaac
     5941 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     6001 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     6061 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     6121 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     6181 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     6241 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca
     6301 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     6361 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     6421 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6481 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     6541 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     6601 cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg
     6661 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     6721 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     6781 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     6841 agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga
     6901 cacacccgcc agcggccgc
//
