LOCUS       sequence-300617-        6639 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 6639)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6639)
  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-m4252892
FEATURES             Location/Qualifiers
     source          1..6639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    151..222
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(227..273)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    291..339
                     /note="synthetic polyadenylation signal"
     enhancer        420..723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        725..927
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     878..898
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      996..1005
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1038..1058
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1059..1100
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1169..1290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1180..1199)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1234..1253
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1405..1451
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1481..1617
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1662..1733
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1959..2547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2039..2058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3047..4256
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4093..4112
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4260..5288
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5315..5363
                     /note="synthetic polyadenylation signal"
     CDS             join(5629..5697,5698..6489)
                     /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(5847..5866)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6502..6638
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
       61 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
      121 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
      181 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
      241 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
      301 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
      361 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      421 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      481 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      541 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      601 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      661 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      721 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      781 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      841 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      901 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg
      961 atcctaccat ccactcgaca cacccgccag cggccgccac catggctgat gcacaatcgt
     1021 ttttaaacgg gtttgcggaa aacctttact ttcagtcggg gaaacctata cccaatccac
     1081 ttctcggctt agattctacc taaggatccg caggcctctg ctagcttgac tgactgagat
     1141 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     1201 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1261 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1321 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     1381 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     1441 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     1501 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     1561 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     1621 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     1681 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     1741 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     1801 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     1861 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     1921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2641 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2761 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     2821 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     2881 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     2941 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     3001 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg
     3061 gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa
     3121 gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
     3181 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
     3241 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
     3301 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
     3361 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
     3421 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
     3481 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
     3541 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
     3601 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
     3661 gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa
     3721 gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca
     3781 ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg
     3841 ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc
     3901 tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag gcttttctcc
     3961 gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc
     4021 tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc
     4081 ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt
     4141 gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt
     4201 gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca
     4261 tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     4321 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     4381 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     4441 atcgttatgt ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca
     4501 ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     4561 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg
     4621 atgccatcgc tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     4681 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     4741 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     4801 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     4861 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     4921 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     4981 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     5041 cccggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     5101 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     5161 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg
     5221 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     5281 aggaataagc tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg
     5341 gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc
     5401 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     5461 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     5521 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     5581 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     5641 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5701 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     5761 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     5821 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5881 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     5941 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6001 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     6061 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6121 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6181 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     6241 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6301 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6361 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6421 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6481 cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     6541 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     6601 ttccatacgt gtcctcctta ccagaaattt atccttaag
//
