LOCUS       sequence-300766-        6642 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 6642)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6642)
  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-m4252896
FEATURES             Location/Qualifiers
     source          1..6642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      243..831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     732..751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1057..1128
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1173..1309)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1339..1385)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1500..1621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1537..1556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1591..1610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1690..1734)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1735..1755)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      1788..1797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1866..2068)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1895..1915)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2070..2373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2454..2502
                     /note="synthetic polyadenylation signal"
     terminator      2520..2566
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2571..2642
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2794..2930)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2943..3734,3735..3803))
                     /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     3566..3585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4069..4117
                     /note="synthetic polyadenylation signal"
     CDS             complement(4144..5172)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(5176..6385)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5320..5339)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ttaaatgcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
       61 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      121 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      181 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      241 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      301 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      361 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      421 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      481 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      541 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      601 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      661 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      721 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      781 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      841 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gatttaaatg
      901 aaactcgtaa tcggagaacg ggtgcaatag caaggaccag atcgaacttg actgacgcaa
      961 ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca agtagctgac ttgactggtt
     1021 aattaagttt cgatagccca aggtaaccaa tgagattagg gagggaagaa agtttgatga
     1081 actacttctg acccctagtg gtgtccagaa aagaccatta aaggaatgtc ttgatcgaaa
     1141 tcgagagatc gcctgcagga tagctgacga ccttaaggat aaatttctgg taaggaggac
     1201 acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg catatgtggg
     1261 agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgcc ggtagatagc
     1321 aatccggccc gaggggcaca aaaaacccct caagacccgt ttagaggccc caaggggtta
     1381 tgctacgata ccgatagaga tgggccaata ccacatttgt agaggtttta cttgctttaa
     1441 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1621 atcatgtctg tgagctgaag gtacgctgta tctcagtcag tcaagctagc agaggcctgc
     1681 ggatccttaa ctgtccatgt gctgcctctc gaacttagct gccgcggttt ctttcgactg
     1741 aaagtaaagg ttttccgcaa acccgtttaa aaacgattgt gcatcagcca tggtggcggc
     1801 cgctggcggg tgtgtcgagt ggatggtagg atcgacaaag atctgatctg acggttcact
     1861 aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     1921 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     1981 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     2041 ccattgatgt actgccaaaa ccgcatcagc atggtaatag cgatgactaa tacgtagatg
     2101 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     2161 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     2221 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     2281 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     2341 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     2401 aatgaccccg taattgatta ctagatctgg ctacactcga tcagcagtta gataataaaa
     2461 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat
     2521 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgaatt gagaggtagc
     2581 tgaagctgct gttatgacca ctagagggaa gaagatacct gtggagctaa tggtccaaga
     2641 tgtcgggagc cgcgatcgct gccaaaaagg taagctgacg atagctgacg agattcgatg
     2701 cccgacgagc aaatgagcgc cacggctgtt cgcgtaagct cgaaacgacg gagattcgat
     2761 agagccagag tcgagtttaa acgattcggg atcttaagga taaatttctg gtaaggagga
     2821 cacgtatgga agtgggcaag ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg
     2881 gagtataaga cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc acctagaatc
     2941 ggttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3181 cagtctatca actgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttacgc
     3241 aaggttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3361 gcggttagct ccttcggtcc tccaatcgtt gtcagaagta agttggccgc agtgttatca
     3421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3481 tctgtgactg gtgaatactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3601 ctcatcattg gaaagcgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3661 tccagttcga tgtaacccac tcgcgcaccc aactgatctt cagcatcttt tactttcacc
     3721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3781 acacggaaat gttgaatact catgcgaaag cctcctgctg atcctgtctc ttgatagatg
     3841 tttgcaaaag ctaggactcg attatacgtt gaccgatcag actatcaata tctaggttta
     3901 aacagatcgg tacgctgccg tgaccaggct tgagtacaat atctgacgat tggacttcct
     3961 gggtcccaag ctggattccg actggctgac tgccaagcaa cgatatggat cgtcgagact
     4021 cgcactgaga ggatcgttct cggtacacca ggcgcgccta gcgttacaca cacaaaaaac
     4081 caacacacat ccatcttcga tggatagcga ttttattatc taagactagg cttacatact
     4141 agcttattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac
     4201 ttctacacag ccatcggtcc agacggcggc gcttctgcgg gcgatttgtg tacgcccgac
     4261 agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc
     4321 gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc
     4381 ccggagccgg ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg
     4441 ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc
     4501 cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac
     4561 attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca
     4621 aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt
     4681 ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta
     4741 ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggcggc
     4801 agcgatggca tccattgcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg
     4861 caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct
     4921 gaactcccca atgtcaagca cttccggaat cgggagcgcg gcggatgcaa agtgccgata
     4981 aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg
     5041 ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc
     5101 ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg
     5161 ctttttaccc atgttgtcta acaaaaaagc caaaaacggc cagaatttag cggacaattt
     5221 acaagtctaa cactgaaaat tacatattga cccaaatgat tacatttcaa aaggtgccta
     5281 aaaaacttca caaaacacac tcgccaaccc cgagcgcata gttcaaaacc ggagcttcag
     5341 ctacttaaga agataggtac ataaaaccga ccaaagaaac tgacgcctca cttatccctc
     5401 ccctcaccag aggtccggcg cctgtcgatt caggagagcc taccctaggc ccgaaccctg
     5461 cgtcctgcga cggagaaaag cctaccgcac acctaccggc tggtggcccc accctgcatt
     5521 ataagccaac agaacgggtg acgtcacgac acgacgaggg cgcgcgctcc caaaggtacg
     5581 ggtgcactgc ccaacggcac cgccataact gccgcccccg caacagacga caaaccgagt
     5641 tctccagtca gtgacaaact tcacgtcagg gtccccagat ggtgcccagc ccatctcacc
     5701 cgaataagag ctttcccgca ttagcgaagg gctcaagacc ttgggttctt gccgcccacc
     5761 atgcccccca ccttgtttca acgacctcac agcccgcctc acaagcgtct tccattcaag
     5821 actcgggaac agccgccatt ttgctgcgct ccccccaacc cccagttcag ggcaaccttg
     5881 ctcgcggacc cagactacag cccttggcgg tctctccaca cgcttccgtc ccaccgagcg
     5941 gcccggcggc cacgaaagcc ccggccagcc cagcagcccg ctactcacca agtgacgatc
     6001 acagcgatcc acaaacaaga accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc
     6061 cacacccggc gcgtccttat ataatcatcg gcgttcaccg ccccacggag atccctccgc
     6121 agaatcgccg agaagggact acttttcctc gcctgttccg ctctctggaa agaaaaccag
     6181 tgccctagag tcacccaagt cccgtcctaa aatgtccttc tgctgatact ggggttctaa
     6241 ggccgagtct tatgagcagc gggccgctgt cctgagcgtc cgggcggaag gatcaggacg
     6301 ctcctgcgcc cttcgtctga cgtggcagcg ctcgccgtga cgaggggggc gcccgcggga
     6361 ggcgccaaaa cccggcgcgg aggccaggcg cgccccgatt tcgatattcc tcggatctcg
     6421 gatagatcgg gattgaggat gtaggttgaa cttctctcgg gtgatcccga tctcggatat
     6481 tcgggattgg agtaatacct ggcgataggc gctgccaagc aatagccaag gcgcaaggcc
     6541 ggccaagaga cttgcgatag ttcgttcgag aggtagcctg caagatggat cggacgaaag
     6601 gtttgaagat tttccgacga gggccttgga gagttcggaa at
//
