LOCUS       sequence-295759-        7112 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 7112)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7112)
  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-m4133692
FEATURES             Location/Qualifiers
     source          1..7112
                     /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..583
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    652..773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(663..682)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     717..736
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      888..934
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      964..1100
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1145..1216
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1442..2030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1522..1541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2530..3739
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3576..3595
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3743..4771
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4798..4846
                     /note="synthetic polyadenylation signal"
     CDS             join(5112..5180,5181..5972)
                     /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(5330..5349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5985..6121
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6266..6337
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6342..6388)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6406..6454
                     /note="synthetic polyadenylation signal"
     enhancer        6535..6838
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6840..7042
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6993..7013
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7111..7112,1..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 agtcgcacca tcaccatcac cattaaggat ccgcaggcct
      601 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
      661 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      721 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
      781 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
      841 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
      901 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
      961 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     1021 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     1081 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     1141 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     1201 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     1261 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     1321 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     1381 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1861 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     2281 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     2341 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     2401 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     2461 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     2521 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     2581 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     2641 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     2701 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     2761 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     2821 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     2881 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     2941 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     3001 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     3061 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     3121 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     3181 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     3241 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     3301 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     3361 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt
     3421 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     3481 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     3541 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     3601 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     3661 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     3721 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     3781 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     3841 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     3901 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     3961 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     4021 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4081 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     4141 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4201 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4261 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4321 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4381 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4441 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4501 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     4561 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4621 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4681 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4741 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     4801 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     4861 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     4921 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     4981 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     5041 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     5101 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5161 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5221 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5281 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5341 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5401 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5461 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5521 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5581 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5641 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     5701 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     5761 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5821 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5881 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5941 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     6001 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     6061 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     6121 agatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     6181 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     6241 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     6301 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     6361 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     6421 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     6481 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     6541 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     6601 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     6661 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     6721 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     6781 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
     6841 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     6901 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     6961 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     7021 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
     7081 accatccact cgacacaccc gccagcggcc gc
//
