LOCUS       sequence-311978-        4620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4620)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4620)
  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-m4134420
FEATURES             Location/Qualifiers
     source          1..4620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             925..945
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             949..1014
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1067..1188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1078..1097)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1132..1151
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1303..1349
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1379..1515
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1560..1631
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1857..2445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1937..1956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3191..3259,3260..4051)
                     /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(3409..3428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4064..4200
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4352..4423
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4428..4474)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4492..4540
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaataa cgagttaagc
      601 ccagtagcgc ttcgccagat gagttgcgcc gcaggtacaa cccaaactgc ttgtacggat
      661 gacaacgccc tcgcttacta taataccaca aagggaggcc gattcgttct ggccctgttg
      721 tcggatctgc aagacttgaa atgggcacgt tttcccaagt ccgacgggac tggcaccatt
      781 tacacagaac tagagccgcc atgcaggttc gtgactgata cccctaaagg acccaaggtc
      841 aaatatctgt attttatcaa ggggcttaac aatctcaata gaggcatggt gttaggatct
      901 ctagccgcaa cggtgcggct gcaagaaaac ctttactttc agtcgatgga ttataaggac
      961 cacgatggag actacaaaga tcatgacatt gattacaagg acgacgatga caaataagct
     1021 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
     1081 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1141 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1201 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1261 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
     1321 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
     1381 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     1441 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     1501 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
     1561 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
     1621 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
     1681 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
     1741 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
     1801 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1861 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1921 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1981 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2041 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2101 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2161 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2221 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2281 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2341 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2401 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2461 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2521 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2581 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2641 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     2701 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     2761 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     2821 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     2881 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     2941 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     3001 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     3061 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     3121 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     3181 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3241 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3301 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3361 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3421 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3481 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     3541 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3601 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3661 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3721 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     3781 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     3841 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3901 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3961 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4021 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     4081 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     4141 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     4201 gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     4261 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     4321 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     4381 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     4441 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     4501 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     4561 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
