LOCUS       sequence-295967-        4776 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 4776)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4776)
  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-m4133488
FEATURES             Location/Qualifiers
     source          1..4776
                     /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"
     CDS             join(2769..2837,2838..3629)
                     /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(2987..3006)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3642..3778
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3930..4001
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4006..4052)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4070..4118
                     /note="synthetic polyadenylation signal"
     enhancer        4199..4502
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4504..4706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4657..4677
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4775..4776,1..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 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2581 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     2641 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     2701 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     2761 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2821 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2881 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2941 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3001 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3061 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3121 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3181 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3241 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3301 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     3361 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3421 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3481 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3541 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3601 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     3661 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     3721 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     3781 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     3841 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     3901 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     3961 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     4021 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     4081 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     4141 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4201 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4261 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4321 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4381 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4441 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4501 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4561 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4621 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4681 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     4741 tcctaccatc cactcgacac acccgccagc ggccgc
//
