LOCUS       sequence-295014-        7133 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 7133)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7133)
  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-m4134454
FEATURES             Location/Qualifiers
     source          1..7133
                     /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..604
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    673..794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(684..703)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     738..757
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      909..955
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      985..1121
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1166..1237
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1463..2051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1543..1562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2551..3760
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3597..3616
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3764..4792
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4819..4867
                     /note="synthetic polyadenylation signal"
     CDS             join(5133..5201,5202..5993)
                     /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(5351..5370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6006..6142
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6287..6358
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6363..6409)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6427..6475
                     /note="synthetic polyadenylation signal"
     enhancer        6556..6859
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6861..7063
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7014..7034
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7132..7133,1..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 cggggaaacc tatacccaat ccacttctcg gcttagattc
      601 tacctaagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
      661 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      721 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      781 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
      841 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
      901 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
      961 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
     1021 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     1081 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
     1141 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
     1201 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
     1261 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
     1321 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
     1381 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1441 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1501 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1561 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1621 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1681 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1741 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1801 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1861 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1921 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1981 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2041 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2101 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2161 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2221 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2281 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     2341 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     2401 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     2461 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     2521 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     2581 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     2641 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     2701 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     2761 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     2821 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     2881 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     2941 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     3001 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     3061 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     3121 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     3181 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     3241 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     3301 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     3361 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3421 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     3481 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     3541 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     3601 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     3661 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     3721 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     3781 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     3841 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     3901 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     3961 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     4021 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     4081 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     4141 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     4201 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     4261 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4321 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4381 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4441 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4501 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     4561 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     4621 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     4681 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     4741 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     4801 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     4861 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     4921 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     4981 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     5041 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     5101 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     5161 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5221 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     5281 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     5341 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5401 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     5461 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5521 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     5581 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5641 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     5701 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     5761 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5821 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5881 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5941 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     6001 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     6061 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     6121 cttaccagaa atttatcctt aagatcgttt aaactcgact ctggctctat cgaatctccg
     6181 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     6241 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     6301 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     6361 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     6421 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     6481 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     6541 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     6601 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     6661 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6721 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6781 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6841 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6901 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6961 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     7021 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     7081 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
