LOCUS       sequence-297317-        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             41..61
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             62..106
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    159..280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(170..189)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     224..243
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      395..441
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      471..607
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    652..723
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(949..1537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1029..1048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2283..2351,2352..3143)
                     /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(2501..2520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3156..3292
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3444..3515
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3520..3566)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3584..3632
                     /note="synthetic polyadenylation signal"
     enhancer        3713..4016
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4018..4220
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4171..4191
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4289..4290,1..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 gaaagaaacc gcggcagcta agttcgagag gcagcacatg gacagttaag ctagcttgac
      121 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      181 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      241 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      301 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      361 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      421 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      481 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      541 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      601 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      661 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      721 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      781 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      841 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
      901 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      961 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1021 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1081 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1141 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1201 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1261 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1321 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1381 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1441 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1741 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1801 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1861 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     1921 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     1981 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     2041 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2101 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2161 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2221 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     2281 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2341 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2401 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2461 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2521 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2581 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2641 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2701 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2761 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2821 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     2881 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     2941 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3001 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3061 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3121 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3181 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3241 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     3301 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     3361 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     3421 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     3481 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     3541 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     3601 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     3661 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3721 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3781 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     3841 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     3901 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     3961 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     4021 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4081 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4141 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4201 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4261 catccactcg acacacccgc cagcggccgc
//
