LOCUS       sequence-295470-        4572 bp    DNA     circular SYN 15-APR-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 4572)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4572)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133286
FEATURES             Location/Qualifiers
     source          1..4572
                     /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..388
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    441..562
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(452..471)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     506..525
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      677..723
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      753..889
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    934..1005
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1231..1819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1311..1330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2565..2633,2634..3425)
                     /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(2783..2802)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3438..3574
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3726..3797
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3802..3848)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3866..3914
                     /note="synthetic polyadenylation signal"
     enhancer        3995..4298
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4300..4502
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4453..4473
                     /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 gtacccctat gacgtaccag attatgcgta agctagcttg actgactgag atacagcgta
      421 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      481 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      541 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      601 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
      661 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      721 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
      781 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
      841 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
      901 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
      961 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     1021 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     1081 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     1141 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1741 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1861 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1981 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2041 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     2101 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     2161 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2221 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2281 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct
     2341 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     2401 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     2461 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     2521 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     2581 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2641 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     2701 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     2761 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2821 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     2881 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2941 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     3001 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3061 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3121 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     3181 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3241 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3301 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3361 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3421 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     3481 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     3541 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     3601 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     3661 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     3721 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     3781 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     3841 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     3901 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     3961 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4021 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4081 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4141 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4201 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4261 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     4321 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4381 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4441 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     4501 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     4561 gccagcggcc gc
//
