LOCUS       sequence-295982-        8062 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8062)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8062)
  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-m4133544
FEATURES             Location/Qualifiers
     source          1..8062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1916..1936
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1937..1978
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2031..2152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2042..2061)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2096..2115
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2267..2313
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2343..2479
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2524..2595
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2821..3409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2901..2920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3909..5118
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4955..4974
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5122..5721
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5122..5141)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5618..5638
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5748..5796
                     /note="synthetic polyadenylation signal"
     CDS             join(6062..6130,6131..6922)
                     /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(6280..6299)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6935..7071
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7216..7287
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7292..7338)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7356..7404
                     /note="synthetic polyadenylation signal"
     enhancer        7485..7788
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7790..7992
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7943..7963
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac actcgctatg tcgataacaa cttctgtggc cctgatggct accctcttga
       61 gtgcattaaa gaccttctag cacgtgctgg taaagcttca tgcactttgt ccgaacaact
      121 ggactttatt gacactaaga ggggtgtata ctgctgccgt gaacatgagc atgaaattgc
      181 ttggtacacg gaacgttctg aaaagagcta tgaattgcag acaccttttg aaattaaatt
      241 ggcaaagaaa tttgacacct tcaatgggga atgtccaaat tttgtatttc ccttaaattc
      301 cataatcaag actattcaac caagggttga aaagaaaaag cttgatggct ttatgggtag
      361 aattcgatct gtctatccag ttgcgtcacc aaatgaatgc aaccaaatgt gcctttcaac
      421 tctcatgaag tgtgatcatt gtggtgaaac ttcatggcag acgggcgatt ttgttaaagc
      481 cacttgcgaa ttttgtggca ctgagaattt gactaaagaa ggtgccacta cttgtggtta
      541 cttaccccaa aatgctgttg ttaaaattta ttgtccagca tgtcacaatt cagaagtagg
      601 acctgagcat agtcttgccg aataccataa tgaatctggc ttgaaaacca ttcttcgtaa
      661 gggtggtcgc actattgcct ttggaggctg tgtgttctct tatgttggtt gccataacaa
      721 gtgtgcctat tgggttccac gtgctagcgc taacataggt tgtaaccata caggtgttgt
      781 tggagaaggt tccgaaggtc ttaatgacaa ccttcttgaa atactccaaa aagagaaagt
      841 caacatcaat attgttggtg actttaaact taatgaagag atcgccatta ttttggcatc
      901 tttttctgct tccacaagtg cttttgtgga aactgtgaaa ggtttggatt ataaagcatt
      961 caaacaaatt gttgaatcct gtggtaattt taaagttaca aaaggaaaag ctaaaaaagg
     1021 tgcctggaat attggtgaac agaaatcaat actgagtcct ctttatgcat ttgcatcaga
     1081 ggctgctcgt gttgtacgat caattttctc ccgcactctt gaaactgctc aaaattctgt
     1141 gcgtgtttta cagaaggccg ctataacaat actagatgga atttcacagt attcactgag
     1201 actcattgat gctatgatgt tcacatctga tttggctact aacaatctag ttgtaatggc
     1261 ctacattaca ggtggtgttg ttcagttgac ttcgcagtgg ctaactaaca tctttggcac
     1321 tgtttatgaa aaactcaaac ccgtccttga ttggcttgaa gagaagttta aggaaggtgt
     1381 agagtttctt agagacggtt gggaaattgt taaatttatc tcaacctgtg cttgtgaaat
     1441 tgtcggtgga caaattgtca cctgtgcaaa ggaaattaag gagagtgttc agacattctt
     1501 taagcttgta aataaatttt tggctttgtg tgctgactct atcattattg gtggagctaa
     1561 acttaaagcc ttgaatttag gtgaaacatt tgtcacgcac tcaaagggat tgtacagaaa
     1621 gtgtgttaaa tccagagaag aaactggcct actcatgcct ctaaaagccc caaaagaaat
     1681 tatcttctta gagggagaaa cacttcccac agaagtgtta acagaggaag ttgtcttgaa
     1741 aactggtgat ttacaaccat tagaacaacc tactagtgaa gctgttgaag ctccattggt
     1801 tggtacacca gtttgtatta acgggcttat gttgctcgaa atcaaagaca cagaaaagta
     1861 ctgtgccctt gcacctaata tgatggtaac aaacaatacc ttcacactca aaggcgaaaa
     1921 cctttacttt cagtcgggga aacctatacc caatccactt ctcggcttag attctaccta
     1981 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
     2041 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2101 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2161 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2221 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     2281 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     2341 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     2401 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     2461 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     2521 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     2581 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     2641 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     2701 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     2761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3241 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     3661 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     3721 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     3781 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     3841 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     3901 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     3961 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     4021 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     4081 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     4141 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     4201 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     4261 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     4321 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     4381 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     4441 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     4501 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     4561 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     4621 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     4681 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     4741 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     4801 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     4861 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     4921 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     4981 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     5041 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     5101 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     5161 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     5221 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5281 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5341 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5401 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
     5461 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5521 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
     5581 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5641 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5701 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
     5761 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     5821 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     5881 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     5941 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     6001 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     6061 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6121 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6181 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6241 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6301 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6361 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6421 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6481 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6541 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6601 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     6661 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6721 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6781 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6841 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6901 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     6961 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     7021 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcgttta
     7081 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     7141 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     7201 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     7261 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     7321 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     7381 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     7441 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     7501 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7561 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     7621 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     7681 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     7741 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     7801 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     7861 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     7921 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     7981 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     8041 cgacacaccc gccagcggcc gc
//
