LOCUS       sequence-300593-        4290 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 4290)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4290)
  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-m4133477
FEATURES             Location/Qualifiers
     source          1..4290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(747..815,816..1607)
                     /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(965..984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1620..1756
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1908..1979
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1984..2030)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2048..2096
                     /note="synthetic polyadenylation signal"
     enhancer        2177..2480
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2482..2684
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2635..2655
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2753..2762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2795..2815
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2816..2860
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    2913..3034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2924..2943)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2978..2997
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3149..3195
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3225..3361
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3406..3477
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(join(3703..4290,1))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3783..3802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
       61 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
      241 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
      301 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
      361 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
      421 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
      481 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
      541 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
      601 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
      661 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
      721 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
      781 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      841 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
      901 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
      961 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1021 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     1081 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1141 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     1201 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1261 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     1321 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     1381 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1441 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1501 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1561 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     1621 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1681 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1741 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     1801 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     1861 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     1921 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     1981 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     2041 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     2101 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     2161 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     2221 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2281 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     2341 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     2401 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     2461 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     2521 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     2581 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     2641 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     2701 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgccaccat
     2761 ggctgatgca caatcgtttt taaacgggtt tgcggaaaac ctttactttc agtcgaaaga
     2821 aaccgcggca gctaagttcg agaggcagca catggacagt taagctagct tgactgactg
     2881 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     2941 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3001 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3061 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3121 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     3181 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     3241 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     3301 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     3361 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     3421 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     3481 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     3541 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     3601 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     3661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4261 gcagattacg cgcagaaaaa aaggatctca
//
