LOCUS       sequence-300355-        7125 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 7125)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7125)
  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-m4134150
FEATURES             Location/Qualifiers
     source          1..7125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    112..183
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(188..234)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    252..300
                     /note="synthetic polyadenylation signal"
     enhancer        381..684
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        686..888
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     839..859
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      957..966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1500..1520
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1521..1547
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1616..1737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1627..1646)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1681..1700
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1852..1898
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1928..2064
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2109..2180
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2406..2994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2486..2505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3494..4703
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4540..4559
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4707..5735
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5762..5810
                     /note="synthetic polyadenylation signal"
     CDS             join(6076..6144,6145..6936)
                     /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(6294..6313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6949..7085
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
       61 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
      121 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
      181 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
      241 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
      301 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
      361 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      421 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      481 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      541 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      601 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      661 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga
      721 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      781 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      841 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      901 cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca
      961 ccatggagag ccttgtccct ggtttcaacg agaaaacaca cgtccaactc agtttgcctg
     1021 ttttacaggt tcgcgacgtg ctcgtacgtg gctttggaga ctccgtggag gaggtcttat
     1081 cagaggcacg tcaacatctt aaagatggca cttgtggctt agtagaagtt gaaaaaggcg
     1141 ttttgcctca acttgaacag ccctatgtgt tcatcaaacg ttcggatgct cgaactgcac
     1201 ctcatggtca tgttatggtt gagctggtag cagaactcga aggcattcag tacggtcgta
     1261 gtggtgagac acttggtgtc cttgtccctc atgtgggcga aataccagtg gcttaccgca
     1321 aggttcttct tcgtaagaac ggtaataaag gagctggtgg ccatagttac ggcgccgatc
     1381 taaagtcatt tgacttaggc gacgagcttg gcactgatcc ttatgaagat tttcaagaaa
     1441 actggaacac taaacatagc agtggtgtta cccgtgaact catgcgtgag cttaacggag
     1501 aaaaccttta ctttcagtcg tacccctatg acgtaccaga ttatgcgtaa ggatccgcag
     1561 gcctctgcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1621 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1681 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1741 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1801 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     1861 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     1921 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     1981 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     2041 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     2101 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     2161 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     2221 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     2281 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     2341 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2401 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2461 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2521 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2581 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2641 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2701 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2761 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2821 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2881 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2941 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3001 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3061 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3121 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3181 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     3241 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     3301 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     3361 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     3421 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     3481 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
     3541 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
     3601 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     3661 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     3721 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     3781 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     3841 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     3901 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     3961 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     4021 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     4081 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     4141 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     4201 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     4261 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     4321 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
     4381 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     4441 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     4501 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     4561 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     4621 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     4681 cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg
     4741 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     4801 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     4861 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg
     4921 cgctcccgat tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca
     4981 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     5041 ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga
     5101 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     5161 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     5221 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     5281 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     5341 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     5401 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     5461 ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc
     5521 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     5581 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     5641 gcagaagcgc cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     5701 gacgccccag cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga
     5761 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     5821 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     5881 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     5941 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     6001 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     6061 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6121 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6181 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6241 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     6301 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6361 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     6421 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6481 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6541 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6601 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     6661 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     6721 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6781 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6841 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6901 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     6961 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     7021 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     7081 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaat
//
