LOCUS       sequence-295936-        4773 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 4773)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4773)
  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-m4133856
FEATURES             Location/Qualifiers
     source          1..4773
                     /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..589
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    642..763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(653..672)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     707..726
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      878..924
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      954..1090
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1135..1206
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1432..2020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1512..1531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2766..2834,2835..3626)
                     /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(2984..3003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3639..3775
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3927..3998
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4003..4049)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4067..4115
                     /note="synthetic polyadenylation signal"
     enhancer        4196..4499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4501..4703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4654..4674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4772..4773,1..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 cgtaccccta tgacgtacca gattatgcgt aagctagctt
      601 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      661 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      721 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      781 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      841 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
      901 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
      961 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     1021 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     1081 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     1141 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     1201 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     1261 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1321 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1861 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2041 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2221 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     2281 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2341 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2401 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2461 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2521 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     2581 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     2641 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     2701 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     2761 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2821 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2881 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2941 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3001 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3061 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3121 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3181 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3241 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3301 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     3361 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     3421 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3481 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3541 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3601 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     3661 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     3721 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     3781 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     3841 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     3901 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     3961 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     4021 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     4081 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     4141 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4201 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4261 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4321 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4381 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4441 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     4501 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4561 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4621 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4681 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     4741 taccatccac tcgacacacc cgccagcggc cgc
//
