LOCUS       sequence-307307-        4269 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 4269)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4269)
  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-m4134024
FEATURES             Location/Qualifiers
     source          1..4269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             47..67
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             68..85
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    138..259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(149..168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     203..222
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      374..420
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      450..586
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    631..702
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(928..1516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1008..1027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2262..2330,2331..3122)
                     /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(2480..2499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3135..3271
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3423..3494
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3499..3545)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3563..3611
                     /note="synthetic polyadenylation signal"
     enhancer        3692..3995
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3997..4199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4150..4170
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgcac catcaccatc accattaagc tagcttgact gactgagata cagcgtacct
      121 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      181 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      241 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
      301 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
      361 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      421 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
      481 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
      541 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
      601 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
      661 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
      721 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
      781 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
      841 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
      901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1741 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     1801 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     1861 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     1921 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     1981 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag
     2041 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     2101 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     2161 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     2221 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     2281 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2341 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     2401 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     2461 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2521 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     2581 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2641 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     2701 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2761 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2821 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     2881 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2941 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3001 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3061 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3121 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     3181 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     3241 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     3301 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     3361 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     3421 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     3481 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     3541 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     3601 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     3661 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     3721 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     3781 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     3841 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     3901 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     3961 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     4021 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4081 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4141 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     4201 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     4261 agcggccgc
//
