LOCUS       sequence-297468-        4263 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 4263)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4263)
  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-m4133855
FEATURES             Location/Qualifiers
     source          1..4263
                     /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             62..79
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    132..253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(143..162)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     197..216
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      368..414
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      444..580
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    625..696
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(922..1510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1002..1021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2256..2324,2325..3116)
                     /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(2474..2493)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3129..3265
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3417..3488
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3493..3539)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3557..3605
                     /note="synthetic polyadenylation signal"
     enhancer        3686..3989
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3991..4193
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4144..4164
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4262..4263,1..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 gcaccatcac catcaccatt aagctagctt gactgactga gatacagcgt accttcagct
      121 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      181 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      241 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
      301 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
      361 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
      421 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
      481 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
      541 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
      601 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
      661 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
      721 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
      781 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
      841 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1741 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     1801 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     1861 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     1921 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     1981 ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     2041 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     2101 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     2161 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     2221 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     2281 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2341 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     2401 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     2461 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2521 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     2581 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2641 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     2701 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2761 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     2821 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     2881 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2941 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3001 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3061 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     3121 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     3181 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     3241 ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat
     3301 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     3361 tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc
     3421 ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc
     3481 tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat
     3541 caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     3601 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
     3661 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3721 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3781 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3841 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     3901 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     3961 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
     4021 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4081 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4141 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     4201 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     4261 cgc
//
