LOCUS       sequence-295061-        5301 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 5301)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5301)
  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-m4134048
FEATURES             Location/Qualifiers
     source          1..5301
                     /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             596..673
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             701..721
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1170..1291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1181..1200)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1235..1254
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1406..1452
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1482..1618
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1663..1734
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1960..2548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2040..2059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3294..3362,3363..4154)
                     /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(3512..3531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4167..4303
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4455..4526
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4531..4577)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4595..4643
                     /note="synthetic polyadenylation signal"
     enhancer        4724..5027
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5029..5231
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5182..5202
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5300..5301,1..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 cgggacggcg aatcccgggg ctgataaatc cctggaaaag
      601 gagatggaag aagaacttta ttgcagtgtc tgcggctaac cggttcaaaa agatcagttc
      661 aagcggcgcc ctagattacg acattcctac gaccgcctcc gagaatctct atttccaggg
      721 tgaatttggc ttggcacaac atgacgaggc cgtcgataac aagtttaata aagaacagca
      781 gaacgctttc tatgagatcc ttcacttacc aaatctgaac gaagagcaac gcaacgcttt
      841 cattcagagc ctgaaagacg atccctcgca gtctgcaaat ctcctggccg aagccaagaa
      901 gctgaatgac gcacaggctc caaaagttga taacaagttt aacaaggagc agcagaatgc
      961 tttctacgag atattgcacc ttcctaatct gaatgaggaa caaagaaatg catttatcca
     1021 gtccctaaaa gacgatccat cacagagtgc caacctgctt gcggaagcca aaaagctcaa
     1081 tgatgcccag gccccgaaag tagacgcgaa ccatcaataa gctagcttga ctgactgaga
     1141 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1201 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1261 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1321 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1381 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1441 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1501 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1561 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1621 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1681 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1741 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1801 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1861 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1921 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1981 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2041 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2101 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2161 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2221 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2281 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2341 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2401 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2461 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2521 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2581 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2641 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2701 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2761 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2821 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2881 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2941 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     3001 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     3061 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     3121 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     3181 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     3241 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     3301 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3361 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     3421 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3481 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3541 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3601 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3661 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     3721 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3781 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3841 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3901 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3961 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4021 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4081 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4141 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     4201 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4261 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     4321 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     4381 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     4441 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     4501 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     4561 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     4621 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     4681 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4741 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4801 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4861 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4921 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4981 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     5041 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     5101 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     5161 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     5221 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     5281 gacacacccg ccagcggccg c
//
