LOCUS       sequence-295075-        4788 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 4788)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4788)
  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-m4134193
FEATURES             Location/Qualifiers
     source          1..4788
                     /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..604
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    657..778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(668..687)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     722..741
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      893..939
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      969..1105
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1150..1221
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1447..2035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1527..1546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2781..2849,2850..3641)
                     /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(2999..3018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3654..3790
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3942..4013
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4018..4064)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4082..4130
                     /note="synthetic polyadenylation signal"
     enhancer        4211..4514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4516..4718
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4669..4689
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4787..4788,1..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 cggggaaacc tatacccaat ccacttctcg gcttagattc
      601 tacctaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
      661 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      721 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      781 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      841 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
      901 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
      961 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     1021 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1081 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1141 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1201 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1261 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1321 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1381 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1441 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1501 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1561 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1621 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1681 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1741 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1801 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1861 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1921 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1981 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2041 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2101 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2161 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2221 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2281 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2341 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2401 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2461 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2521 atcggggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     2581 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     2641 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     2701 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     2761 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     2821 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2881 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2941 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     3001 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3061 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     3121 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3181 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3241 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3301 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     3361 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     3421 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3481 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3541 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3601 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     3661 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     3721 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     3781 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     3841 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     3901 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     3961 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     4021 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     4081 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     4141 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     4201 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4261 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4321 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4381 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4441 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4501 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4561 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4621 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4681 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     4741 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
