LOCUS       sequence-295912-        5136 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 5136)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5136)
  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-m4133824
FEATURES             Location/Qualifiers
     source          1..5136
                     /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             911..928
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1005..1126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1016..1035)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1070..1089
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1241..1287
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1317..1453
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1498..1569
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1795..2383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1875..1894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3129..3197,3198..3989)
                     /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(3347..3366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4002..4138
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4290..4361
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4366..4412)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4430..4478
                     /note="synthetic polyadenylation signal"
     enhancer        4559..4862
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4864..5066
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5017..5037
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5135..5136,1..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 cgatgagtga caaaatcatt catctaaccg atgactcctt
      601 tgatacagac gtgttgaagg ccgatggtgc gatactggta gatttctggg ctgaatggtg
      661 tggcccgtgc aagatgatcg cacccattct ggacgagatc gccgacgagt atcaaggaaa
      721 attaactgtc gctaagctta atatagatca gaacccaggg acggcaccta aatacggcat
      781 caggggaata cctaccctcc tgctctttaa gaatggggaa gttgccgcga caaaagtggg
      841 agccctgtct aagggccagc taaaagagtt cttggacgct aacctggcag gttcaggctc
      901 ggggcacatg caccatcacc atcaccatag cagcgggctc gtgccacgcg gctaagctag
      961 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     1021 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1081 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1141 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1201 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     1261 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     1321 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     1381 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     1441 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     1501 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1561 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1621 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     1681 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1741 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1801 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1861 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1921 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1981 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2041 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2101 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2161 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2221 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2281 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2341 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2401 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2461 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2521 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2581 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     2641 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     2701 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2761 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2821 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2881 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2941 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     3001 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     3061 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     3121 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3241 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3301 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3421 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3541 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3661 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     3721 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3961 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     4021 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     4081 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     4141 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     4201 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     4261 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     4321 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     4381 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     4441 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     4501 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4561 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4621 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4681 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4741 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4801 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4861 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4921 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4981 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     5041 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     5101 tcctaccatc cactcgacac acccgccagc ggccgc
//
