LOCUS       sequence-295374-        5139 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 5139)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5139)
  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-m4134120
FEATURES             Location/Qualifiers
     source          1..5139
                     /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             914..931
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1008..1129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1019..1038)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1073..1092
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1244..1290
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1320..1456
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1501..1572
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1798..2386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1878..1897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3132..3200,3201..3992)
                     /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(3350..3369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4005..4141
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4293..4364
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4369..4415)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4433..4481
                     /note="synthetic polyadenylation signal"
     enhancer        4562..4865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4867..5069
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5020..5040
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5138..5139,1..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 agtcgatgag tgacaaaatc attcatctaa ccgatgactc
      601 ctttgataca gacgtgttga aggccgatgg tgcgatactg gtagatttct gggctgaatg
      661 gtgtggcccg tgcaagatga tcgcacccat tctggacgag atcgccgacg agtatcaagg
      721 aaaattaact gtcgctaagc ttaatataga tcagaaccca gggacggcac ctaaatacgg
      781 catcagggga atacctaccc tcctgctctt taagaatggg gaagttgccg cgacaaaagt
      841 gggagccctg tctaagggcc agctaaaaga gttcttggac gctaacctgg caggttcagg
      901 ctcggggcac atgcaccatc accatcacca tagcagcggg ctcgtgccac gcggctaagc
      961 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
     1021 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1081 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1141 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1201 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1261 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1321 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1381 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1441 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     1501 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1561 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1621 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1681 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2221 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2521 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2581 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2641 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2701 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2761 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2821 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2881 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2941 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     3001 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     3061 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     3121 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3241 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3301 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3421 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3541 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3661 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3721 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3961 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     4021 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     4081 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     4141 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     4201 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     4261 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     4321 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     4381 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     4441 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     4501 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4561 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4621 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4681 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4741 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4801 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4861 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4921 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4981 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     5041 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     5101 cgatcctacc atccactcga cacacccgcc agcggccgc
//
