LOCUS       sequence-295891-        6568 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 6568)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6568)
  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-m4134327
FEATURES             Location/Qualifiers
     source          1..6568
                     /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             440..484
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    537..658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(548..567)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     602..621
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      773..819
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      849..985
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1030..1101
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1327..1915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1407..1426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2415..3624
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3461..3480
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3628..4227
                     /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(3628..3647)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4124..4144
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4254..4302
                     /note="synthetic polyadenylation signal"
     CDS             join(4568..4636,4637..5428)
                     /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(4786..4805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5441..5577
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5722..5793
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5798..5844)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5862..5910
                     /note="synthetic polyadenylation signal"
     enhancer        5991..6294
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6296..6498
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6449..6469
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6567..6568,1..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 aagaaaccgc ggcagctaag ttcgagaggc agcacatgga
      481 cagttaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
      541 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      601 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      661 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      721 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
      781 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
      841 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
      901 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
      961 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1021 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1081 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1141 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1201 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1261 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1741 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2161 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2221 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2281 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2341 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2401 atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt
     2461 cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg
     2521 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     2581 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     2641 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     2701 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     2761 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     2821 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     2881 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     2941 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     3001 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     3061 gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca
     3121 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     3181 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     3241 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
     3301 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     3361 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     3421 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     3481 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     3541 tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg
     3601 ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc
     3661 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     3721 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     3781 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     3841 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     3901 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc
     3961 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4021 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc
     4081 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4141 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     4201 tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa
     4261 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc
     4321 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     4381 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     4441 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     4501 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     4561 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4621 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4681 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4741 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4801 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4861 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4921 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4981 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5041 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5101 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     5161 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     5221 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5281 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5341 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5401 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     5461 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     5521 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     5581 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     5641 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     5701 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     5761 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     5821 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     5881 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     5941 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6001 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6061 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6121 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     6181 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6241 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
     6301 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     6361 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     6421 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     6481 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
     6541 tccactcgac acacccgcca gcggccgc
//
