LOCUS       sequence-295665-        7052 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 7052)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7052)
  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-m4134019
FEATURES             Location/Qualifiers
     source          1..7052
                     /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..523
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    592..713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(603..622)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     657..676
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      828..874
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      904..1040
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1085..1156
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1382..1970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1462..1481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2470..3679
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3516..3535
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3683..4711
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4738..4786
                     /note="synthetic polyadenylation signal"
     CDS             join(5052..5120,5121..5912)
                     /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(5270..5289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5925..6061
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6206..6277
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6282..6328)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6346..6394
                     /note="synthetic polyadenylation signal"
     enhancer        6475..6778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6780..6982
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6933..6953
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7051..7052,1..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 agtcgaagag gcgatggaaa aagaatttta tcgcggtgag
      481 tgcagccaac agattcaaaa agatatcctc gtctggtgcc ctctaaggat ccgcaggcct
      541 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
      601 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      661 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
      721 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
      781 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
      841 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
      901 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
      961 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     1021 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     1081 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     1141 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     1201 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     1261 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     1321 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1561 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1801 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1981 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2041 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2101 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     2221 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     2281 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     2341 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     2401 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     2461 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     2521 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     2581 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     2641 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     2701 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     2761 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     2821 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     2881 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     2941 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     3001 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     3061 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     3121 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     3181 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     3241 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     3301 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt
     3361 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     3421 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     3481 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     3541 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     3601 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     3661 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     3721 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     3781 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     3841 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     3901 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     3961 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4021 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     4081 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4141 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4201 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4261 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4321 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4381 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4441 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     4501 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4561 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4621 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4681 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     4741 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     4801 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     4861 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     4921 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     4981 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     5041 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5101 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5161 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5221 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5281 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5341 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5401 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5461 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5521 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5581 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     5641 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     5701 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5761 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5821 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5881 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     5941 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     6001 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     6061 agatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     6121 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     6181 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     6241 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     6301 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     6361 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     6421 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     6481 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     6541 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     6601 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     6661 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     6721 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
     6781 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     6841 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     6901 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     6961 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
     7021 accatccact cgacacaccc gccagcggcc gc
//
