LOCUS       sequence-297453-        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"
     CDS             47..67
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             101..178
                     /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             206..226
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    691..812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(702..721)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     756..775
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      927..973
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1003..1139
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1184..1255
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1481..2069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1561..1580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2569..3778
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3615..3634
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3782..4810
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4837..4885
                     /note="synthetic polyadenylation signal"
     CDS             join(5151..5219,5220..6011)
                     /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(5369..5388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6024..6160
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6312..6383
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6388..6434)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6452..6500
                     /note="synthetic polyadenylation signal"
     enhancer        6581..6884
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6886..7088
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7039..7059
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcggga cggcgaatcc cggggctgat aaatccctgg aaaaggagat ggaagaagaa
      121 ctttattgca gtgtctgcgg ctaaccggtt caaaaagatc agttcaagcg gcgccctaga
      181 ttacgacatt cctacgaccg cctccgagaa tctctatttc cagggtgaat ttggcttggc
      241 acaacatgac gaggccgtcg ataacaagtt taataaagaa cagcagaacg ctttctatga
      301 gatccttcac ttaccaaatc tgaacgaaga gcaacgcaac gctttcattc agagcctgaa
      361 agacgatccc tcgcagtctg caaatctcct ggccgaagcc aagaagctga atgacgcaca
      421 ggctccaaaa gttgataaca agtttaacaa ggagcagcag aatgctttct acgagatatt
      481 gcaccttcct aatctgaatg aggaacaaag aaatgcattt atccagtccc taaaagacga
      541 tccatcacag agtgccaacc tgcttgcgga agccaaaaag ctcaatgatg cccaggcccc
      601 gaaagtagac gcgaaccatc aataaggatc cgcaggcctc tgctagcttg actgactgag
      661 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
      721 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
      781 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
      841 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
      901 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
      961 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
     1021 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     1081 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     1141 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
     1201 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
     1261 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
     1321 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
     1381 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
     1441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1921 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2281 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
     2341 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
     2401 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
     2461 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
     2521 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc
     2581 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg
     2641 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     2701 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     2761 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     2821 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     2881 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     2941 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     3001 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     3061 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     3121 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     3181 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga
     3241 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt
     3301 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     3361 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     3421 tctgttggct tataatgcag ggtggggcca ccagccggta ggtgtgcggt aggcttttct
     3481 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     3541 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     3601 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     3661 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     3721 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa
     3781 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     3841 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     3901 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     3961 agatcgttat gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga
     4021 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     4081 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat
     4141 ggatgccatc gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     4201 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     4261 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     4321 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     4381 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4441 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     4501 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     4561 gccccggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     4621 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     4681 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga
     4741 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     4801 aaaggaataa gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga
     4861 tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga
     4921 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     4981 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     5041 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     5101 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     5161 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5221 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     5281 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     5341 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5401 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     5461 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5521 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     5581 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5641 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5701 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5761 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5821 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5881 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5941 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6001 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     6061 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     6121 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     6181 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     6241 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     6301 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     6361 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     6421 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     6481 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     6541 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6601 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     6661 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6721 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     6781 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     6841 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     6901 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     6961 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     7021 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     7081 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     7141 acacccgcca gcggccgc
//
