LOCUS       sequence-295833-        7043 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 7043)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7043)
  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-m4134426
FEATURES             Location/Qualifiers
     source          1..7043
                     /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             449..514
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    583..704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(594..613)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     648..667
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      819..865
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      895..1031
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1076..1147
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1373..1961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1453..1472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2461..3670
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3507..3526
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3674..4702
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4729..4777
                     /note="synthetic polyadenylation signal"
     CDS             join(5043..5111,5112..5903)
                     /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(5261..5280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5916..6052
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6197..6268
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6273..6319)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6337..6385
                     /note="synthetic polyadenylation signal"
     enhancer        6466..6769
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6771..6973
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6924..6944
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7042..7043,1..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 agtcgatgga ttataaggac cacgatggag actacaaaga
      481 tcatgacatt gattacaagg acgacgatga caaataagga tccgcaggcc tctgctagct
      541 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
      601 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
      661 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
      721 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
      781 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
      841 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
      901 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
      961 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     1021 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
     1081 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
     1141 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
     1201 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     1261 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     1321 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1381 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1441 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1501 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1561 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1621 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1681 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1741 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1801 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1861 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1921 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1981 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2041 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2101 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2161 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     2221 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     2281 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     2341 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     2401 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     2461 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg
     2521 ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc
     2581 ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg
     2641 acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga
     2701 aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg
     2761 attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc
     2821 gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg
     2881 gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc
     2941 aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc
     3001 agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag
     3061 gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc
     3121 gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga
     3181 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     3241 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     3301 gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg
     3361 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     3421 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     3481 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     3541 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
     3601 atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt
     3661 tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     3721 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     3781 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     3841 tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc tcccgattcc
     3901 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     3961 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     4021 cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg ggttcggccc
     4081 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     4141 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     4201 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     4261 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     4321 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     4381 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4441 gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg accaactcta
     4501 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     4561 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgccgc
     4621 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     4681 tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa taaaatcgct
     4741 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg
     4801 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     4861 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     4921 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     4981 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     5041 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5101 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5161 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5221 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5281 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5341 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5401 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5461 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5521 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5581 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     5641 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     5701 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5761 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5821 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5881 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     5941 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     6001 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt
     6061 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     6121 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     6181 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     6241 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     6301 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     6361 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     6421 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6481 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6541 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6601 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6661 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6721 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6781 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6841 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6901 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6961 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     7021 tcgacacacc cgccagcggc cgc
//
