LOCUS       sequence-342384-        8514 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 8514)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8514)
  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-m4133966
FEATURES             Location/Qualifiers
     source          1..8514
                     /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             2494..2514
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2515..2556
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2625..2746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2636..2655)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2690..2709
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2861..2907
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2937..3073
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3118..3189
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3415..4003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3495..3514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4503..5712
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5549..5568
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5716..6744
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6771..6819
                     /note="synthetic polyadenylation signal"
     CDS             join(7085..7153,7154..7945)
                     /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(7303..7322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7958..8094
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8246..8317
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8322..8368)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8386..8434
                     /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 ccatgtacac tcgctatgtc
      601 gataacaact tctgtggccc tgatggctac cctcttgagt gcattaaaga ccttctagca
      661 cgtgctggta aagcttcatg cactttgtcc gaacaactgg actttattga cactaagagg
      721 ggtgtatact gctgccgtga acatgagcat gaaattgctt ggtacacgga acgttctgaa
      781 aagagctatg aattgcagac accttttgaa attaaattgg caaagaaatt tgacaccttc
      841 aatggggaat gtccaaattt tgtatttccc ttaaattcca taatcaagac tattcaacca
      901 agggttgaaa agaaaaagct tgatggcttt atgggtagaa ttcgatctgt ctatccagtt
      961 gcgtcaccaa atgaatgcaa ccaaatgtgc ctttcaactc tcatgaagtg tgatcattgt
     1021 ggtgaaactt catggcagac gggcgatttt gttaaagcca cttgcgaatt ttgtggcact
     1081 gagaatttga ctaaagaagg tgccactact tgtggttact taccccaaaa tgctgttgtt
     1141 aaaatttatt gtccagcatg tcacaattca gaagtaggac ctgagcatag tcttgccgaa
     1201 taccataatg aatctggctt gaaaaccatt cttcgtaagg gtggtcgcac tattgccttt
     1261 ggaggctgtg tgttctctta tgttggttgc cataacaagt gtgcctattg ggttccacgt
     1321 gctagcgcta acataggttg taaccataca ggtgttgttg gagaaggttc cgaaggtctt
     1381 aatgacaacc ttcttgaaat actccaaaaa gagaaagtca acatcaatat tgttggtgac
     1441 tttaaactta atgaagagat cgccattatt ttggcatctt tttctgcttc cacaagtgct
     1501 tttgtggaaa ctgtgaaagg tttggattat aaagcattca aacaaattgt tgaatcctgt
     1561 ggtaatttta aagttacaaa aggaaaagct aaaaaaggtg cctggaatat tggtgaacag
     1621 aaatcaatac tgagtcctct ttatgcattt gcatcagagg ctgctcgtgt tgtacgatca
     1681 attttctccc gcactcttga aactgctcaa aattctgtgc gtgttttaca gaaggccgct
     1741 ataacaatac tagatggaat ttcacagtat tcactgagac tcattgatgc tatgatgttc
     1801 acatctgatt tggctactaa caatctagtt gtaatggcct acattacagg tggtgttgtt
     1861 cagttgactt cgcagtggct aactaacatc tttggcactg tttatgaaaa actcaaaccc
     1921 gtccttgatt ggcttgaaga gaagtttaag gaaggtgtag agtttcttag agacggttgg
     1981 gaaattgtta aatttatctc aacctgtgct tgtgaaattg tcggtggaca aattgtcacc
     2041 tgtgcaaagg aaattaagga gagtgttcag acattcttta agcttgtaaa taaatttttg
     2101 gctttgtgtg ctgactctat cattattggt ggagctaaac ttaaagcctt gaatttaggt
     2161 gaaacatttg tcacgcactc aaagggattg tacagaaagt gtgttaaatc cagagaagaa
     2221 actggcctac tcatgcctct aaaagcccca aaagaaatta tcttcttaga gggagaaaca
     2281 cttcccacag aagtgttaac agaggaagtt gtcttgaaaa ctggtgattt acaaccatta
     2341 gaacaaccta ctagtgaagc tgttgaagct ccattggttg gtacaccagt ttgtattaac
     2401 gggcttatgt tgctcgaaat caaagacaca gaaaagtact gtgcccttgc acctaatatg
     2461 atggtaacaa acaatacctt cacactcaaa ggcgaaaacc tttactttca gtcggggaaa
     2521 cctataccca atccacttct cggcttagat tctacctaag gatccgcagg cctctgctag
     2581 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     2641 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2701 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2761 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     2821 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     2881 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     2941 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     3001 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     3061 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     3121 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     3181 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     3241 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     3301 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     3361 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3421 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3481 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3541 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3601 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3661 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3721 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3781 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3841 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3901 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3961 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4021 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4081 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4141 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4201 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     4261 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     4321 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     4381 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     4441 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     4501 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     4561 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     4621 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     4681 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     4741 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     4801 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     4861 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     4921 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     4981 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     5041 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     5101 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     5161 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     5221 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     5281 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     5341 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
     5401 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     5461 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     5521 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     5581 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     5641 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     5701 tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt
     5761 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
     5821 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
     5881 gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc gctcccgatt
     5941 ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt
     6001 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     6061 gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag cgggttcggc
     6121 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     6181 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     6241 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     6301 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     6361 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     6421 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     6481 gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     6541 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     6601 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc
     6661 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     6721 actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat aataaaatcg
     6781 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg
     6841 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc
     6901 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc
     6961 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac
     7021 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt
     7081 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7141 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7201 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7261 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7321 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7381 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7441 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7501 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7561 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7621 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct
     7681 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct
     7741 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7801 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7861 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7921 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc
     7981 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     8041 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc
     8101 gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     8161 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     8221 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     8281 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     8341 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     8401 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     8461 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
