LOCUS       sequence-306184-        7128 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 7128)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7128)
  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-m4133516
FEATURES             Location/Qualifiers
     source          1..7128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      171..307
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    459..530
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(535..581)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    599..647
                     /note="synthetic polyadenylation signal"
     enhancer        728..1031
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1033..1235
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1186..1206
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1304..1313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1850..1870
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1871..1897
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1966..2087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1977..1996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2031..2050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2202..2248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2278..2414
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2459..2530
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2756..3344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2836..2855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3844..5053
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4890..4909
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5057..6085
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6112..6160
                     /note="synthetic polyadenylation signal"
     CDS             join(6426..6494,6495..7128,1..158)
                     /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(6644..6663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
       61 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
      121 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
      181 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      241 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      301 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
      361 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
      421 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
      481 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
      541 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
      601 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
      661 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
      721 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      781 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      841 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      901 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      961 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1021 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     1081 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     1141 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     1201 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     1261 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgccacca tggagtctct
     1321 cgttcccggc tttaatgaaa aaactcatgt ccagttatcc ttgccagtgc ttcaagtaag
     1381 ggatgtgctg gtgcggggtt tcggagacag tgtagaggaa gtcctgtcag aggctagaca
     1441 gcacctgaag gatgggacct gtggcttagt ggaagttgag aagggagtcc tcccgcagct
     1501 ggaacaacct tatgtgttta taaaacgcag cgacgcgcga acagcacctc acgggcatgt
     1561 aatggtggag cttgtcgccg agttggaggg catccagtac ggaagatcgg gtgaaacgct
     1621 gggagttcta gtgccccacg ttggcgaaat tccggtggcc taccgtaagg ttttgctcag
     1681 gaaaaacgga aataaggggg ctggtggcca tagctatggg gcagacctga agtctttcga
     1741 tctgggcgac gagctgggca ccgacccata tgaggatttc caggaaaact ggaacacaaa
     1801 acacagctcc ggggtaactc gcgagctaat gcgagaattg aatggagggg aaaaccttta
     1861 ctttcagtcg tacccctatg acgtaccaga ttatgcgtaa ggatccgcag gcctctgcta
     1921 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
     1981 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2041 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2101 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     2161 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
     2221 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
     2281 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
     2341 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
     2401 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
     2461 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
     2521 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
     2581 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     2641 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     2701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3181 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3361 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     3601 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     3661 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     3721 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     3781 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     3841 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     3901 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     3961 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     4021 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     4081 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     4141 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     4201 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     4261 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     4321 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     4381 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     4441 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     4501 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     4561 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     4621 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     4681 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt
     4741 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     4801 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     4861 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     4921 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     4981 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     5041 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     5101 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     5161 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     5221 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat
     5281 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     5341 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     5401 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg
     5461 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     5521 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     5581 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     5641 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     5701 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     5761 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     5821 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     5881 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     5941 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     6001 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     6061 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc
     6121 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     6181 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     6241 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     6301 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     6361 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     6421 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6481 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6541 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6601 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6661 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6721 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6781 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6841 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6901 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6961 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     7021 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     7081 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggag
//
