LOCUS       sequence-295699-        6676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6676)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6676)
  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-m4133440
FEATURES             Location/Qualifiers
     source          1..6676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             545..565
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             566..592
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    645..766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(656..675)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     710..729
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      881..927
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      957..1093
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1138..1209
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1435..2023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1515..1534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2523..3732
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3569..3588
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3736..4335
                     /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(3736..3755)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4232..4252
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4362..4410
                     /note="synthetic polyadenylation signal"
     CDS             join(4676..4744,4745..5536)
                     /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(4894..4913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5549..5685
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5830..5901
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5906..5952)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5970..6018
                     /note="synthetic polyadenylation signal"
     enhancer        6099..6402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6404..6606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6557..6577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6675..6676,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggag tctctcgttc ccggctttaa tgaaaaaact catgtccagt tatccttgcc
       61 agtgcttcaa gtaagggatg tgctggtgcg gggtttcgga gacagtgtag aggaagtcct
      121 gtcagaggct agacagcacc tgaaggatgg gacctgtggc ttagtggaag ttgagaaggg
      181 agtcctcccg cagctggaac aaccttatgt gtttataaaa cgcagcgacg cgcgaacagc
      241 acctcacggg catgtaatgg tggagcttgt cgccgagttg gagggcatcc agtacggaag
      301 atcgggtgaa acgctgggag ttctagtgcc ccacgttggc gaaattccgg tggcctaccg
      361 taaggttttg ctcaggaaaa acggaaataa gggggctggt ggccatagct atggggcaga
      421 cctgaagtct ttcgatctgg gcgacgagct gggcaccgac ccatatgagg atttccagga
      481 aaactggaac acaaaacaca gctccggggt aactcgcgag ctaatgcgag aattgaatgg
      541 aggggaaaac ctttactttc agtcgtaccc ctatgacgta ccagattatg cgtaagctag
      601 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      661 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      721 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      781 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      841 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
      901 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
      961 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     1021 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     1081 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     1141 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1201 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1261 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     1321 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1381 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1441 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1501 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1561 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1621 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1681 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1741 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1801 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1861 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1921 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1981 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2041 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2101 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2161 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2221 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     2281 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     2341 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2401 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2461 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2521 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2581 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2641 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     2701 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     2761 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     2821 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     2881 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     2941 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     3001 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     3061 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     3121 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     3181 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     3241 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     3301 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     3361 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
     3421 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3481 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3541 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3601 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3661 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     3721 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3781 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3841 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3901 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3961 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4021 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     4081 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4141 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     4201 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4261 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4321 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     4381 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     4441 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4501 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4561 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4621 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4681 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4741 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     4801 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4861 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4921 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4981 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5041 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     5101 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5161 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5221 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     5281 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5341 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5401 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5461 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5521 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5581 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5641 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcg tttaaactcg
     5701 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt
     5761 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc
     5821 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa
     5881 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa
     5941 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg
     6001 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt
     6061 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     6301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     6361 acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag
     6421 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     6481 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     6541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     6601 agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac
     6661 acccgccagc ggccgc
//
