LOCUS       sequence-295360-        7019 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 7019)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7019)
  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-m4134117
FEATURES             Location/Qualifiers
     source          1..7019
                     /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..490
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    559..680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(570..589)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     624..643
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      795..841
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      871..1007
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1052..1123
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1349..1937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1429..1448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2437..3646
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3483..3502
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3650..4678
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4705..4753
                     /note="synthetic polyadenylation signal"
     CDS             join(5019..5087,5088..5879)
                     /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(5237..5256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5892..6028
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6173..6244
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6249..6295)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6313..6361
                     /note="synthetic polyadenylation signal"
     enhancer        6442..6745
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6747..6949
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6900..6920
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7018..7019,1..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 agtcgaaaga aaccgcggca gctaagttcg agaggcagca
      481 catggacagt taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
      541 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
      601 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      661 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
      721 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
      781 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      841 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
      901 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
      961 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
     1021 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     1081 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     1141 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     1201 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     1261 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     1321 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1381 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1441 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1501 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1561 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1621 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1681 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1741 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1801 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1861 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2041 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2101 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2161 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     2221 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     2281 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     2341 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     2401 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     2461 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     2521 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     2581 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     2641 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     2701 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     2761 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     2821 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     2881 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     2941 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     3001 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     3061 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     3121 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     3181 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     3241 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     3301 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     3361 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     3421 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     3481 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     3541 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     3601 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     3661 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     3721 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3781 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3841 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     3901 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     3961 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     4021 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4081 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4141 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4201 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4261 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4321 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4381 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     4441 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4501 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4561 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     4621 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     4681 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     4741 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     4801 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     4861 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     4921 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     4981 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     5041 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5101 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     5161 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     5221 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5281 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     5341 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5401 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     5461 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5521 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     5581 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     5641 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5701 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5761 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5821 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5881 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     5941 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     6001 gtcctcctta ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa
     6061 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     6121 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     6181 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     6241 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     6301 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     6361 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     6421 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6481 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6541 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6601 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6661 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6721 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     6781 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6841 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6901 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     6961 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
