LOCUS       sequence-297573-        6137 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 6137)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6137)
  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-m4252974
FEATURES             Location/Qualifiers
     source          1..6137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    99..220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(110..129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     164..183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      335..381
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      411..547
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    592..663
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(889..1477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(969..988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1977..3186
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3023..3042
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3190..3789
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3190..3209)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3686..3706
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3816..3864
                     /note="synthetic polyadenylation signal"
     CDS             join(4130..4198,4199..4990)
                     /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(4348..4367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5003..5139
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5291..5362
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5367..5413)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5431..5479
                     /note="synthetic polyadenylation signal"
     enhancer        5560..5863
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5865..6067
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6018..6038
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtgtaag ctagcttgac
       61 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      121 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      181 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      241 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      301 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      361 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      421 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      481 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      541 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      601 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      661 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      721 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      781 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
      841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1081 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1321 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1441 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1561 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1621 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1681 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1741 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1801 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     1861 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     1921 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     1981 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2041 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2101 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2161 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2221 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2281 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2341 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2401 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2461 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2521 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     2581 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     2641 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     2701 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     2761 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     2821 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     2881 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     2941 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3001 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3061 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3121 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3181 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3241 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3301 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3361 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3421 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3481 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3541 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3601 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     3661 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3721 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     3781 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     3841 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     3901 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     3961 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4021 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4081 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4141 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4201 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4261 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4321 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4381 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4441 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4501 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4561 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4621 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4681 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4741 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4801 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4861 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4921 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4981 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5041 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5101 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5161 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5221 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     5281 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     5341 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     5401 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     5461 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5521 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5581 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5641 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5701 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5761 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     5821 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     5881 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     5941 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6001 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6061 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6121 cacccgccag cggccgc
//
