LOCUS       sequence-294991-        4767 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 4767)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4767)
  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-m4133382
FEATURES             Location/Qualifiers
     source          1..4767
                     /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"
     CDS             join(2760..2828,2829..3620)
                     /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(2978..2997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3633..3769
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3921..3992
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3997..4043)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4061..4109
                     /note="synthetic polyadenylation signal"
     enhancer        4190..4493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4495..4697
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4648..4668
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4766..4767,1..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 cctggtgtac
     2521 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     2581 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     2641 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     2701 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     2761 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2821 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     2881 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2941 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3001 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3061 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3121 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     3181 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3241 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3301 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     3361 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3421 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3481 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3541 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3601 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     3661 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     3721 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     3781 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     3841 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     3901 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     3961 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     4021 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     4081 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     4141 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4201 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4261 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4321 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4381 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4441 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     4501 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4561 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4621 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4681 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     4741 ccactcgaca cacccgccag cggccgc
//
