LOCUS       sequence-317515-        7008 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 7008)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7008)
  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-m4134407
FEATURES             Location/Qualifiers
     source          1..7008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1003..1023
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1024..1050
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1119..1240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1130..1149)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1184..1203
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1355..1401
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1431..1567
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1612..1683
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1909..2497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1989..2008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2997..4206
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4043..4062
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4210..5238
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5265..5313
                     /note="synthetic polyadenylation signal"
     CDS             join(5579..5647,5648..6439)
                     /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(5797..5816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6452..6588
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6740..6811
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6816..6862)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6880..6928
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggcagg gaatgcgaca
      601 gaggttcctg ccaactcaac ggtcttgtct ttttgtgctt tcgccgtgga cgctgcaaaa
      661 gcctacaagg attatctagc ctcgggtgga cagccaatca ctaactgcgt aaagatgctt
      721 tgtacccaca ccggcactgg ccaagcaatt acagtgaccc ccgaagctaa tatggaccag
      781 gagtccttcg gaggtgcgag ctgctgcctg tactgtcgtt gtcatataga tcacccgaat
      841 cccaaagggt tttgcgacct caagggcaaa tatgtccaga tcccaacgac atgcgccaac
      901 gatcctgtgg gattcactct gaagaacacc gtatgtaccg tttgcgggat gtggaagggg
      961 tatggctgta gttgcgacca attacgggaa ccgatgctgc aagaaaacct ttactttcag
     1021 tcgtacccct atgacgtacc agattatgcg taaggatccg caggcctctg ctagcttgac
     1081 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     1141 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1201 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1261 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1321 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     1381 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     1441 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     1501 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     1561 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     1621 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     1681 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1741 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1801 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2101 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2341 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2521 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2581 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2641 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2701 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     2761 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2821 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2881 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2941 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     3001 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     3061 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     3121 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     3181 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     3241 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     3301 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     3361 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     3421 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     3481 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     3541 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     3601 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     3661 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     3721 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     3781 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     3841 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     3901 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3961 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     4021 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     4081 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     4141 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     4201 ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     4261 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     4321 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     4381 ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc gattccggaa
     4441 gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     4501 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     4561 gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt cggcccattc
     4621 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     4681 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     4741 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     4801 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     4861 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     4921 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     4981 gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     5041 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     5101 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc
     5161 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     5221 ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     5281 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     5341 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     5401 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     5461 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     5521 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     5581 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5641 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     5701 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5761 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5821 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5881 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5941 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     6001 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6061 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6121 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     6181 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6241 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6301 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6361 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6421 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     6481 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     6541 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     6601 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     6661 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     6721 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     6781 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     6841 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     6901 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     6961 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
