LOCUS       sequence-295871-        4791 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 4791)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4791)
  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-m4133666
FEATURES             Location/Qualifiers
     source          1..4791
                     /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             563..607
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    660..781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(671..690)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     725..744
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      896..942
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      972..1108
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1153..1224
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1450..2038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1530..1549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2784..2852,2853..3644)
                     /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(3002..3021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3657..3793
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3945..4016
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4021..4067)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4085..4133
                     /note="synthetic polyadenylation signal"
     enhancer        4214..4517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4519..4721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4672..4692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4790..4791,1..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 cgaaagaaac cgcggcagct aagttcgaga ggcagcacat
      601 ggacagttaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
      661 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      721 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      781 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
      841 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
      901 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
      961 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1021 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1081 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1141 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1201 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1261 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     1321 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     1381 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1861 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1981 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     2281 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     2341 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     2401 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2461 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     2521 gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     2581 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     2641 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     2701 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     2761 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     2821 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2881 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     2941 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     3001 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3061 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     3121 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3181 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     3241 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3301 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     3361 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     3421 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3481 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3541 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3601 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     3661 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     3721 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     3781 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     3841 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     3901 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     3961 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
     4021 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
     4081 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
     4141 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
     4201 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4261 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4321 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4381 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4441 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4501 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4561 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4621 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4681 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     4741 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
