LOCUS       sequence-307382-        7152 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 7152)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7152)
  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-m4258319
FEATURES             Location/Qualifiers
     source          1..7152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      648..657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             690..710
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             744..821
                     /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             849..869
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1334..1455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1345..1364)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1399..1418
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1570..1616
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1646..1782
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1827..1898
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2124..2712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2204..2223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3212..4421
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4258..4277
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4425..5453
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5480..5528
                     /note="synthetic polyadenylation signal"
     CDS             join(5794..5862,5863..6654)
                     /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(6012..6031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6667..6803
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6955..7026
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7031..7077)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7095..7143
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgcc accatggctg
      661 atgcacaatc gtttttaaac gggtttgcgg aaaaccttta ctttcagtcg ggacggcgaa
      721 tcccggggct gataaatccc tggaaaagga gatggaagaa gaactttatt gcagtgtctg
      781 cggctaaccg gttcaaaaag atcagttcaa gcggcgccct agattacgac attcctacga
      841 ccgcctccga gaatctctat ttccagggtg aatttggctt ggcacaacat gacgaggccg
      901 tcgataacaa gtttaataaa gaacagcaga acgctttcta tgagatcctt cacttaccaa
      961 atctgaacga agagcaacgc aacgctttca ttcagagcct gaaagacgat ccctcgcagt
     1021 ctgcaaatct cctggccgaa gccaagaagc tgaatgacgc acaggctcca aaagttgata
     1081 acaagtttaa caaggagcag cagaatgctt tctacgagat attgcacctt cctaatctga
     1141 atgaggaaca aagaaatgca tttatccagt ccctaaaaga cgatccatca cagagtgcca
     1201 acctgcttgc ggaagccaaa aagctcaatg atgcccaggc cccgaaagta gacgcgaacc
     1261 atcaataagg atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag
     1321 ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     1381 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1441 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1501 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta
     1561 tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc
     1621 cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct
     1681 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     1741 ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc
     1801 aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag
     1861 gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct
     1921 atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
     1981 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
     2041 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2101 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2161 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2221 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2281 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2341 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2401 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2461 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2521 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2581 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2641 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2701 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2761 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2821 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2881 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2941 atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt
     3001 cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc
     3061 ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat
     3121 ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag
     3181 atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc
     3241 gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc
     3301 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
     3361 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
     3421 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
     3481 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
     3541 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
     3601 gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
     3661 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
     3721 gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
     3781 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
     3841 ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg
     3901 agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc
     3961 ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta
     4021 cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg
     4081 cagggtgggg ccaccagccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg
     4141 gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg
     4201 gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga
     4261 agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg
     4321 caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat
     4381 tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg
     4441 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     4501 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     4561 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     4621 ggcactttgc atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg
     4681 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     4741 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg
     4801 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     4861 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     4921 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     4981 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     5041 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     5101 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     5161 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt
     5221 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     5281 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     5341 ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac
     5401 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta
     5461 tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt
     5521 tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc
     5581 tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga
     5641 agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa
     5701 cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc
     5761 tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc
     5821 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5881 aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc
     5941 aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact
     6001 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6061 ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag
     6121 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6181 aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt
     6241 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6301 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt
     6361 aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg
     6421 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6481 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6541 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6601 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt
     6661 ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     6721 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     6781 ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg
     6841 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     6901 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     6961 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     7021 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     7081 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     7141 attatctaac tg
//
