LOCUS       sequence-307397-        6999 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 6999)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6999)
  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-m4133343
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     259..278
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             426..1454
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    1481..1529
                     /note="synthetic polyadenylation signal"
     CDS             join(1795..1863,1864..2655)
                     /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(2013..2032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2668..2804
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2956..3027
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3032..3078)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3096..3144
                     /note="synthetic polyadenylation signal"
     enhancer        3225..3528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3530..3732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3683..3703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3801..3810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4227..4247
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4248..4265
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4334..4455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4345..4364)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4399..4418
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4570..4616
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4646..4782
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4827..4898
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5124..5712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5204..5223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6212..6999,1..422)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
       61 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
      121 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
      181 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
      241 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
      301 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
      361 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
      421 acaacatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
      481 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
      541 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
      601 acaaagatcg ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc
      661 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
      721 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
      781 caatggatgc catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac
      841 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
      901 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
      961 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     1021 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     1081 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     1141 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     1201 gatcgccccg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     1261 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     1321 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga
     1381 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     1441 gggcaaagga ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg
     1501 aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga
     1561 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     1621 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     1681 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     1741 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     1801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     1921 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1981 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2041 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2101 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2161 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     2221 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2281 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2341 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     2401 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2461 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2521 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2581 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2641 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     2701 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     2761 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta
     2821 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     2881 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     2941 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     3001 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     3061 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     3121 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     3181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     3541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     3601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     3661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     3721 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     3781 cgacacaccc gccagcggcc gccaccatgg cagggaatgc gacagaggtt cctgccaact
     3841 caacggtctt gtctttttgt gctttcgccg tggacgctgc aaaagcctac aaggattatc
     3901 tagcctcggg tggacagcca atcactaact gcgtaaagat gctttgtacc cacaccggca
     3961 ctggccaagc aattacagtg acccccgaag ctaatatgga ccaggagtcc ttcggaggtg
     4021 cgagctgctg cctgtactgt cgttgtcata tagatcaccc gaatcccaaa gggttttgcg
     4081 acctcaaggg caaatatgtc cagatcccaa cgacatgcgc caacgatcct gtgggattca
     4141 ctctgaagaa caccgtatgt accgtttgcg ggatgtggaa ggggtatggc tgtagttgcg
     4201 accaattacg ggaaccgatg ctgcaagaaa acctttactt tcagtcgcac catcaccatc
     4261 accattaagg atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag
     4321 ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4381 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4441 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4501 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta
     4561 tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc
     4621 cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct
     4681 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     4741 ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc
     4801 aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag
     4861 gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct
     4921 atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
     4981 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
     5041 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5101 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5161 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5221 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5281 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5341 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5401 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5461 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5521 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5581 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5641 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5701 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5761 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5821 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5881 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5941 atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt
     6001 cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc
     6061 ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat
     6121 ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag
     6181 atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc
     6241 gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc
     6301 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
     6361 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
     6421 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
     6481 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
     6541 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
     6601 gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
     6661 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
     6721 gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
     6781 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
     6841 ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg
     6901 agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc
     6961 ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtg
//
