LOCUS       sequence-295461-        4647 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 4647)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4647)
  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-m4134138
FEATURES             Location/Qualifiers
     source          1..4647
                     /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"
     CDS             join(2640..2708,2709..3500)
                     /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(2858..2877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3513..3649
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3801..3872
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3877..3923)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3941..3989
                     /note="synthetic polyadenylation signal"
     enhancer        4070..4373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4375..4577
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4528..4548
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4646..4647,1..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 cctggtgtac
     2401 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     2461 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     2521 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     2581 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     2641 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2701 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     2761 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2821 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2881 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2941 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3001 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     3061 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3121 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3181 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     3241 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3301 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3361 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3421 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3481 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     3541 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     3601 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     3661 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     3721 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     3781 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     3841 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     3901 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     3961 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     4021 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4081 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4141 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4201 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4261 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4321 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     4381 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4441 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4501 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4561 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     4621 ccactcgaca cacccgccag cggccgc
//
