LOCUS       sequence-307305-        6542 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 6542)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6542)
  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-m4252792
FEATURES             Location/Qualifiers
     source          1..6542
                     /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    504..625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(515..534)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     569..588
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      740..786
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      816..952
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    997..1068
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1294..1882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1374..1393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2382..3591
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3428..3447
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3595..4194
                     /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(3595..3614)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4091..4111
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4221..4269
                     /note="synthetic polyadenylation signal"
     CDS             join(4535..4603,4604..5395)
                     /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(4753..4772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5408..5544
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5696..5767
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5772..5818)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5836..5884
                     /note="synthetic polyadenylation signal"
     enhancer        5965..6268
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6270..6472
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6423..6443
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6541..6542,1..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 ctaagctagc ttgactgact gagatacagc
      481 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
      541 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
      601 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
      661 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
      721 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
      781 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
      841 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
      901 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
      961 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1021 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1081 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1141 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1201 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2101 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2161 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2221 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2281 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     2341 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     2401 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     2461 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     2521 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     2581 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     2641 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     2701 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     2761 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     2821 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     2881 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     2941 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     3001 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     3061 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     3121 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     3181 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     3241 gcttataatg cagggtgggg ccaccagccg gtaggtgtgc ggtaggcttt tctccgtcgc
     3301 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     3361 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     3421 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     3481 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     3541 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc
     3601 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3661 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3721 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3781 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3841 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3901 ctggccgcgc agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg
     3961 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     4021 gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc
     4081 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4141 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt
     4201 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     4261 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     4321 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     4381 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     4441 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     4501 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     4561 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4621 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     4681 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     4741 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4801 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     4861 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4921 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     4981 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5041 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     5101 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     5161 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5221 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5281 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5341 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     5401 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     5461 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     5521 tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc tggctctatc
     5581 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     5641 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     5701 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     5761 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     5821 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     5881 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     5941 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6001 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     6061 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     6121 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6181 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6241 ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca tcaatgggcg
     6301 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     6361 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     6421 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     6481 gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc gccagcggcc
     6541 gc
//
