LOCUS       sequence-295454-        6571 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 6571)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6571)
  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-m4134124
FEATURES             Location/Qualifiers
     source          1..6571
                     /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..487
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    540..661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(551..570)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     605..624
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      776..822
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      852..988
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1033..1104
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1330..1918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1410..1429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2418..3627
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3464..3483
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3631..4230
                     /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(3631..3650)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4127..4147
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4257..4305
                     /note="synthetic polyadenylation signal"
     CDS             join(4571..4639,4640..5431)
                     /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(4789..4808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5444..5580
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5725..5796
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5801..5847)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5865..5913
                     /note="synthetic polyadenylation signal"
     enhancer        5994..6297
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6299..6501
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6452..6472
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6570..6571,1..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 agtcggggaa acctataccc aatccacttc tcggcttaga
      481 ttctacctaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
      541 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      601 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      661 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
      721 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
      781 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
      841 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
      901 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
      961 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1021 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1081 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1141 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     1201 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     1261 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1741 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     2161 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     2221 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     2281 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2341 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     2401 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     2461 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     2521 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     2581 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     2641 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     2701 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     2761 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     2821 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     2881 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     2941 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     3001 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     3061 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     3121 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     3181 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     3241 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     3301 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     3361 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     3421 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     3481 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     3541 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     3601 tggccgtttt tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc
     3661 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     3721 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     3781 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     3841 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     3901 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag
     3961 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4021 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc
     4081 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4141 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4201 tggtgcatga cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata
     4261 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc
     4321 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     4381 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     4441 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     4501 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     4561 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4681 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4741 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4801 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4861 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     4921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4981 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5101 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     5161 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     5221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5401 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     5461 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     5521 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     5581 gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca
     5641 gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt
     5701 tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc
     5761 ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc
     5821 gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac
     5881 acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag
     5941 atctagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6001 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6061 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6121 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6181 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg
     6301 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     6361 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     6421 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     6481 gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta
     6541 ccatccactc gacacacccg ccagcggccg c
//
