LOCUS       sequence-295923-        4815 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 4815)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4815)
  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-m4133789
FEATURES             Location/Qualifiers
     source          1..4815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             542..562
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             566..631
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    684..805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(695..714)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     749..768
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      920..966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      996..1132
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1177..1248
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1474..2062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1554..1573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2808..2876,2877..3668)
                     /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(3026..3045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3681..3817
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3969..4040
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4045..4091)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4109..4157
                     /note="synthetic polyadenylation signal"
     enhancer        4238..4541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4543..4745
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4696..4716
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4814..4815,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggag agccttgtcc ctggtttcaa cgagaaaaca cacgtccaac tcagtttgcc
       61 tgttttacag gttcgcgacg tgctcgtacg tggctttgga gactccgtgg aggaggtctt
      121 atcagaggca cgtcaacatc ttaaagatgg cacttgtggc ttagtagaag ttgaaaaagg
      181 cgttttgcct caacttgaac agccctatgt gttcatcaaa cgttcggatg ctcgaactgc
      241 acctcatggt catgttatgg ttgagctggt agcagaactc gaaggcattc agtacggtcg
      301 tagtggtgag acacttggtg tccttgtccc tcatgtgggc gaaataccag tggcttaccg
      361 caaggttctt cttcgtaaga acggtaataa aggagctggt ggccatagtt acggcgccga
      421 tctaaagtca tttgacttag gcgacgagct tggcactgat ccttatgaag attttcaaga
      481 aaactggaac actaaacata gcagtggtgt tacccgtgaa ctcatgcgtg agcttaacgg
      541 agaaaacctt tactttcagt cgatggatta taaggaccac gatggagact acaaagatca
      601 tgacattgat tacaaggacg acgatgacaa ataagctagc ttgactgact gagatacagc
      661 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
      721 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
      781 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
      841 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
      901 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
      961 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1021 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1081 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1141 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1201 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1261 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1321 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1381 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2041 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2101 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2161 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2221 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2281 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2341 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2401 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2461 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     2521 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     2581 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     2641 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     2701 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     2761 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     2821 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2881 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     2941 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     3001 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3061 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     3121 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3181 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     3241 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3301 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3361 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     3421 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3481 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3541 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3601 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3661 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     3721 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     3781 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     3841 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     3901 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     3961 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     4021 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     4081 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     4141 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     4201 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4261 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4321 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4381 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4441 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     4501 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     4561 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4621 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     4681 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     4741 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     4801 cccgccagcg gccgc
//
