LOCUS       sequence-299533-        6648 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 6648)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6648)
  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-m4258332
FEATURES             Location/Qualifiers
     source          1..6648
                     /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             68..112
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    181..302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(192..211)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     246..265
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      417..463
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      493..629
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    674..745
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(971..1559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1051..1070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2059..3268
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3105..3124
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3272..4300
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4327..4375
                     /note="synthetic polyadenylation signal"
     CDS             join(4641..4709,4710..5501)
                     /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(4859..4878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5514..5650
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5802..5873
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5878..5924)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5942..5990
                     /note="synthetic polyadenylation signal"
     enhancer        6071..6374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6376..6578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6529..6549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgaaa gaaaccgcgg cagctaagtt cgagaggcag cacatggaca gttaaggatc
      121 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
      181 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      241 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      301 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      361 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
      421 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
      481 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
      541 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
      601 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
      661 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
      721 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
      781 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
      841 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
      901 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      961 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1021 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1081 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1141 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1201 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1261 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1321 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1381 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1441 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1501 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1561 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1621 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1681 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1741 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     1801 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     1861 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     1921 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     1981 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     2041 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     2101 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     2161 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     2221 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     2281 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     2341 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     2401 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     2461 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     2521 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     2581 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     2641 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     2701 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     2761 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     2821 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     2881 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     2941 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     3001 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     3061 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     3121 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     3181 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     3241 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     3301 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     3361 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     3421 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     3481 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     3541 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     3601 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     3661 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     3721 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     3781 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     3841 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     3901 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     3961 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4021 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     4081 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4141 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     4201 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     4261 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     4321 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     4381 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     4441 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     4501 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     4561 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     4621 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     4681 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4741 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4801 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     4861 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4921 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     4981 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5041 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5101 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5161 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     5221 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     5281 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5341 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5401 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5461 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     5521 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     5581 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     5641 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     5701 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     5761 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     5821 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     5881 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     5941 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     6001 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     6061 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6361 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     6421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     6481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     6541 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     6601 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
