LOCUS       sequence-295221-        4590 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 4590)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4590)
  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-m4133663
FEATURES             Location/Qualifiers
     source          1..4590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             341..361
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             362..406
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    459..580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(470..489)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     524..543
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      695..741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      771..907
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    952..1023
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1249..1837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1329..1348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2583..2651,2652..3443)
                     /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(2801..2820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3456..3592
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3744..3815
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3820..3866)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3884..3932
                     /note="synthetic polyadenylation signal"
     enhancer        4013..4316
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4318..4520
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4471..4491
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaat gagcttagtc ctgttgcact acgacagatg tcttgtgctg ccggtactac
       61 acaaactgct tgcactgatg acaatgcgtt agcttactac aacacaacaa agggaggtag
      121 gtttgtactt gcactgttat ccgatttaca ggatttgaaa tgggctagat tccctaagag
      181 tgatggaact ggtactatct atacagaact ggaaccacct tgtaggtttg ttacagacac
      241 acctaaaggt cctaaagtga agtatttata ctttattaaa ggattaaaca acctaaatag
      301 aggtatggta cttggtagtt tagctgccac agtacgtcta gaaaaccttt actttcagtc
      361 gaaagaaacc gcggcagcta agttcgagag gcagcacatg gacagttaag ctagcttgac
      421 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      481 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      541 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      601 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      661 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      721 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      781 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      841 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      901 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      961 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     1021 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1081 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1141 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1201 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1261 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1321 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1381 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1441 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1501 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1561 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1621 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1681 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1741 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1801 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1861 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1921 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1981 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2041 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     2101 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2161 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2221 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2281 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     2341 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2401 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2461 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2521 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     2581 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2641 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2701 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2761 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2821 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2881 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2941 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3001 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3061 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3121 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     3181 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     3241 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3301 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3361 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3421 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3481 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3541 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     3601 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     3661 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     3721 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     3781 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     3841 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     3901 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     3961 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4021 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4081 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4141 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4201 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4261 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     4321 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4381 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4441 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4501 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4561 catccactcg acacacccgc cagcggccgc
//
