LOCUS       sequence-301662-        7158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7158)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7158)
  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-m4252941
FEATURES             Location/Qualifiers
     source          1..7158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        533..1742
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1579..1598
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1746..2774
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    2801..2849
                     /note="synthetic polyadenylation signal"
     CDS             join(3115..3183,3184..3975)
                     /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(3333..3352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3988..4124
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4276..4347
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4352..4398)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4416..4464
                     /note="synthetic polyadenylation signal"
     enhancer        4545..4848
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4850..5052
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5003..5023
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5169..5189
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5223..5300
                     /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"
     CDS             5328..5348
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    5813..5934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5824..5843)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5878..5897
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      6049..6095
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      6125..6261
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6306..6377
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(join(6603..7158,1..33))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6683..6702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
       61 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      241 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
      301 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
      361 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
      421 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
      481 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg
      541 cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca
      601 gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
      661 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
      721 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
      781 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
      841 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
      901 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
      961 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
     1021 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
     1081 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
     1141 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc
     1201 gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt
     1261 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     1321 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     1381 ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt
     1441 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     1501 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     1561 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     1621 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     1681 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     1741 acaacatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     1801 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     1861 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     1921 acaaagatcg ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc
     1981 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     2041 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     2101 caatggatgc catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac
     2161 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     2221 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     2281 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     2341 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     2401 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     2461 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     2521 gatcgccccg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     2581 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     2641 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga
     2701 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     2761 gggcaaagga ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg
     2821 aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga
     2881 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     2941 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     3001 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     3061 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     3121 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3181 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     3241 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3301 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3361 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3421 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3481 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     3541 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3601 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3661 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     3721 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3781 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3841 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3901 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3961 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     4021 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     4081 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta
     4141 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     4201 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     4261 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     4321 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     4381 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     4441 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     4501 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     4561 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     4621 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     4681 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     4741 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     4801 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     4861 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     4921 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     4981 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     5041 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     5101 cgacacaccc gccagcggcc gccaccatga gtgccgatgc tcagtccttt ctcaacggtt
     5161 tcgcagtgga aaacctttac tttcagtcgg gacggcgaat cccggggctg ataaatccct
     5221 ggaaaaggag atggaagaag aactttattg cagtgtctgc ggctaaccgg ttcaaaaaga
     5281 tcagttcaag cggcgcccta gattacgaca ttcctacgac cgcctccgag aatctctatt
     5341 tccagggtga atttggcttg gcacaacatg acgaggccgt cgataacaag tttaataaag
     5401 aacagcagaa cgctttctat gagatccttc acttaccaaa tctgaacgaa gagcaacgca
     5461 acgctttcat tcagagcctg aaagacgatc cctcgcagtc tgcaaatctc ctggccgaag
     5521 ccaagaagct gaatgacgca caggctccaa aagttgataa caagtttaac aaggagcagc
     5581 agaatgcttt ctacgagata ttgcaccttc ctaatctgaa tgaggaacaa agaaatgcat
     5641 ttatccagtc cctaaaagac gatccatcac agagtgccaa cctgcttgcg gaagccaaaa
     5701 agctcaatga tgcccaggcc ccgaaagtag acgcgaacca tcaataagga tccgcaggcc
     5761 tctgctagct tgactgactg agatacagcg taccttcagc tcacagacat gataagatac
     5821 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5881 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     5941 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     6001 aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc
     6061 ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct
     6121 accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     6181 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     6241 ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat
     6301 caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa
     6361 actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt
     6421 caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc
     6481 agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa
     6541 atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7021 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7141 ggtttttttg tttgcaag
//
