LOCUS       sequence-300328-        7106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7106)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7106)
  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-m4133639
FEATURES             Location/Qualifiers
     source          1..7106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    62..133
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(138..184)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    202..250
                     /note="synthetic polyadenylation signal"
     enhancer        331..634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        636..838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     789..809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1834..1854
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1858..1923
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1976..2097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1987..2006)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2041..2060
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2212..2258
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2288..2424
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2469..2540
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2766..3354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2846..2865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3854..5063
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4900..4919
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5067..5666
                     /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(5067..5086)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5563..5583
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5693..5741
                     /note="synthetic polyadenylation signal"
     CDS             join(6007..6075,6076..6867)
                     /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(6225..6244)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6880..7016
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
       61 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
      121 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
      181 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
      241 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
      301 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      481 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      541 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      601 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
      661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      781 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      841 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
      901 gcggccgcca ccatgagtgg gttccggaaa atggcgtttc cctccggtaa ggtcgaagga
      961 tgcatggttc aggtgacctg tggcacaact acgttaaacg gcctatggct ggacgatgtg
     1021 gtatactgtc ctcgacacgt catctgcaca tcagaggaca tgctgaatcc aaactatgag
     1081 gatttgctca taagaaagag caatcataac tttctggtgc aagctggaaa tgtacagctt
     1141 agggttattg ggcactctat gcagaactgc gtgttaaaac tcaaggtgga taccgccaat
     1201 ccgaaaacac ctaagtacaa attcgtgcgc atccagccag gccaaacctt ctctgtcctg
     1261 gcatgttata acggcagccc ctcaggtgtc taccagtgcg ccatgcgccc taattttact
     1321 attaagggtt ccttcttgaa cgggtcgtgc ggatcggtcg gctttaatat cgactatgac
     1381 tgtgtgagtt tttgctacat gcatcacatg gaacttccga ctggggttca cgctggaacg
     1441 gatctggagg gtaacttcta tggacccttc gtggatagac agaccgccca ggcagccggc
     1501 actgacacta ccattacggt gaatgttcta gcttggctct acgcagctgt aataaacggc
     1561 gatcgttggt tcttgaacag gtttacaaca accctgaacg actttaatct agtcgccatg
     1621 aaatataatt atgaaccact gacccaagat catgtagaca ttctcgggcc actgagcgca
     1681 cagacaggga tcgctgtgct ggacatgtgt gccagtttaa aggagctgct tcagaatggt
     1741 atgaacggcc gaacaatcct agggtctgcg ctcttagagg atgaatttac ccccttcgat
     1801 gtagtgcggc aatgtagcgg cgttactttt caagaaaacc tttactttca gtcgatggat
     1861 tataaggacc acgatggaga ctacaaagat catgacattg attacaagga cgacgatgac
     1921 aaataagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1981 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2041 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2101 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2161 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     2221 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     2281 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     2341 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     2401 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     2461 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     2521 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     2581 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     2641 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     2701 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2761 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2821 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2881 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2941 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3001 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3061 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3121 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3181 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3241 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3301 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3361 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3421 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3481 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3541 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     3601 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     3661 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     3721 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     3781 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     3841 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
     3901 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
     3961 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     4021 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     4081 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     4141 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     4201 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     4261 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     4321 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     4381 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     4441 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     4501 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     4561 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     4621 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     4681 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
     4741 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     4801 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     4861 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     4921 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     4981 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     5041 cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg cgcctcgcca
     5101 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5161 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5221 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5281 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     5341 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggagggcc
     5401 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5461 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg
     5521 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5581 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5641 gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag ataataaaat
     5701 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct
     5761 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     5821 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     5881 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     5941 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     6001 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6061 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6121 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6181 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6241 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6301 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6361 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6421 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6481 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6541 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     6601 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     6661 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6721 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6781 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6841 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     6901 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     6961 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     7021 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     7081 acagccgtgg cgctcatttg ctcgtc
//
