LOCUS       sequence-295362-        4794 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 4794)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4794)
  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-m4133545
FEATURES             Location/Qualifiers
     source          1..4794
                     /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..610
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    663..784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(674..693)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     728..747
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      899..945
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      975..1111
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1156..1227
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1453..2041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1533..1552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2787..2855,2856..3647)
                     /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(3005..3024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3660..3796
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3948..4019
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4024..4070)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4088..4136
                     /note="synthetic polyadenylation signal"
     enhancer        4217..4520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4522..4724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4675..4695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4793..4794,1..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 agtcgaaaga aaccgcggca gctaagttcg agaggcagca
      601 catggacagt taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
      661 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      721 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      781 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
      841 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
      901 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
      961 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     1021 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     1081 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     1141 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     1201 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     1261 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     1321 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1381 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2101 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2161 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2221 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     2281 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     2341 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2401 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2461 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2521 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     2581 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     2641 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     2701 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     2761 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     2821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2881 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     2941 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     3001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3061 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     3121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3181 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     3241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     3361 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     3421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     3661 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     3721 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     3781 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     3841 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     3901 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     3961 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     4021 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     4081 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     4141 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     4201 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4261 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4321 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4381 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4441 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4501 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     4561 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4621 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4681 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     4741 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
