LOCUS       sequence-295734-        6667 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 6667)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6667)
  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-m4133758
FEATURES             Location/Qualifiers
     source          1..6667
                     /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..583
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    636..757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(647..666)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     701..720
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      872..918
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      948..1084
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1129..1200
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1426..2014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1506..1525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2514..3723
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3560..3579
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3727..4326
                     /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(3727..3746)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4223..4243
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4353..4401
                     /note="synthetic polyadenylation signal"
     CDS             join(4667..4735,4736..5527)
                     /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(4885..4904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5540..5676
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5821..5892
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5897..5943)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5961..6009
                     /note="synthetic polyadenylation signal"
     enhancer        6090..6393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6395..6597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6548..6568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6666..6667,1..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 agtcgcacca tcaccatcac cattaagcta gcttgactga
      601 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
      661 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      721 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      781 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      841 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
      901 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
      961 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     1021 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     1081 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     1141 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     1201 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     1261 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     1321 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     1381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1441 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1501 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1561 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1621 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1681 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1741 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1801 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1861 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1921 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1981 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2041 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2101 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2161 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2221 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     2281 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     2341 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     2401 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     2461 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     2521 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     2581 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     2641 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     2701 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     2761 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     2821 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     2881 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     2941 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     3001 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     3061 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     3121 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     3181 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     3241 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     3301 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     3361 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     3421 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     3481 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     3541 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     3601 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     3661 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     3721 gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3781 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     3841 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     3901 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     3961 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4021 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
     4081 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4141 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
     4201 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4261 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4321 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     4381 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     4441 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     4501 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     4561 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     4621 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     4681 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4741 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     4801 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     4861 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4921 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     4981 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5041 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     5101 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5161 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5221 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     5281 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5341 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5401 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5461 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5521 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     5581 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     5641 ccatacgtgt cctccttacc agaaatttat ccttaagatc gtttaaactc gactctggct
     5701 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     5761 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     5821 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     5881 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     5941 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     6001 gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca
     6061 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     6121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     6181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     6241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     6301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6361 tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat
     6421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     6481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     6541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     6601 ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag
     6661 cggccgc
//
