LOCUS       sequence-300562-        6675 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 6675)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6675)
  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-m4252876
FEATURES             Location/Qualifiers
     source          1..6675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    165..236
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(241..287)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    305..353
                     /note="synthetic polyadenylation signal"
     enhancer        434..737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        739..941
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     892..912
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1010..1019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1052..1072
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1073..1150
                     /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"
     polyA_signal    1219..1340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1230..1249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1284..1303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1455..1501
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1531..1667
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1712..1783
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2009..2597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2089..2108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3097..4306
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4143..4162
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4310..5338
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5365..5413
                     /note="synthetic polyadenylation signal"
     CDS             join(5679..5747,5748..6539)
                     /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(5897..5916)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      join(6552..6675,1..13)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
       61 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
      121 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
      181 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
      241 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
      301 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
      361 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
      421 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      481 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      541 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      601 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      661 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      721 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      781 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      841 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      901 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      961 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg ccaccatggc
     1021 tgatgcacaa tcgtttttaa acgggtttgc ggaaaacctt tactttcagt cgaagaggcg
     1081 atggaaaaag aattttatcg cggtgagtgc agccaacaga ttcaaaaaga tatcctcgtc
     1141 tggtgccctc taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
     1201 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1261 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1321 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1381 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
     1441 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1501 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
     1561 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     1621 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
     1681 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     1741 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     1801 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     1861 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     1921 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     1981 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2041 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2101 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2161 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2221 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2281 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2341 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2401 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2461 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2521 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2581 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2641 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2701 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2821 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     2881 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     2941 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     3001 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     3061 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     3121 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     3181 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     3241 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     3301 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     3361 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     3421 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     3481 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     3541 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     3601 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     3661 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     3721 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     3781 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     3841 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     3901 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     3961 taatgcaggg tggggccacc agccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     4021 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     4081 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     4141 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     4201 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     4261 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     4321 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4381 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4441 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4501 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     4561 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4621 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     4681 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4741 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4801 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4861 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4921 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4981 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     5041 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     5101 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     5161 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     5221 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     5281 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     5341 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     5401 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     5461 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     5521 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     5581 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     5641 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     5701 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5761 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     5821 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     5881 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5941 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     6001 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6061 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     6121 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6181 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     6241 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     6301 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6361 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6421 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6481 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6541 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     6601 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     6661 gtcctcctta ccaga
//
