LOCUS       sequence-297463-        6987 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 6987)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6987)
  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-m4252897
FEATURES             Location/Qualifiers
     source          1..6987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..61
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             410..427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    520..641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(531..550)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     585..604
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      756..802
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      832..968
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1013..1084
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1310..1898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1390..1409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2398..3607
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3444..3463
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3611..4639
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4666..4714
                     /note="synthetic polyadenylation signal"
     CDS             join(4980..5048,5049..5840)
                     /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(5198..5217)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5853..5989
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6141..6212
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6217..6263)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6281..6329
                     /note="synthetic polyadenylation signal"
     enhancer        6410..6713
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6715..6917
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6868..6888
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6986..6987,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gatgagtgac aaaatcattc atctaaccga tgactccttt gatacagacg tgttgaaggc
      121 cgatggtgcg atactggtag atttctgggc tgaatggtgt ggcccgtgca agatgatcgc
      181 acccattctg gacgagatcg ccgacgagta tcaaggaaaa ttaactgtcg ctaagcttaa
      241 tatagatcag aacccaggga cggcacctaa atacggcatc aggggaatac ctaccctcct
      301 gctctttaag aatggggaag ttgccgcgac aaaagtggga gccctgtcta agggccagct
      361 aaaagagttc ttggacgcta acctggcagg ttcaggctcg gggcacatgc accatcacca
      421 tcaccatagc agcgggctcg tgccacgcgg ctaaggatcc gcaggcctct gctagcttga
      481 ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg
      541 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      601 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
      661 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
      721 acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa
      781 acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg
      841 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
      901 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
      961 atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt
     1021 taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc
     1081 cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact
     1141 tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat
     1201 ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa
     1261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1741 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1921 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1981 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2041 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2101 cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc
     2161 cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg
     2221 aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg
     2281 ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct
     2341 acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc
     2401 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca
     2461 cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     2521 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     2581 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     2641 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     2701 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     2761 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     2821 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     2881 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     2941 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     3001 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct
     3061 aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt
     3121 gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg
     3181 cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac
     3241 gtcacccgtt ctgttggctt ataatgcagg gtggggccac cagccggtag gtgtgcggta
     3301 ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca
     3361 ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt
     3421 atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc
     3481 gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg
     3541 taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt
     3601 gttagacaac atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3661 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3721 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     3781 tttctacaaa gatcgttatg tttatcggca ctttgcatcc gccgcgctcc cgattccgga
     3841 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     3901 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     3961 ggaggcaatg gatgccatcg ctgccgccga tcttagccag acgagcgggt tcggcccatt
     4021 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4081 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4141 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4201 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4261 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4321 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4381 tgcaggatcg ccccggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4441 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4501 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgccgccgt
     4561 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4621 tccgagggca aaggaataag ctagtatgta agcctagtct tagataataa aatcgctatc
     4681 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac
     4741 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     4801 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     4861 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     4921 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     4981 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5041 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     5101 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5161 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5221 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5281 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5341 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     5401 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5461 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5521 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     5581 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5641 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5701 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5761 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5821 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     5881 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     5941 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     6001 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     6061 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     6121 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     6181 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     6241 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     6301 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     6361 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     6421 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     6481 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     6541 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     6601 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     6661 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     6721 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     6781 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     6841 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     6901 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     6961 ccactcgaca cacccgccag cggccgc
//
