LOCUS       sequence-295806-        7163 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 7163)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7163)
  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-m4133814
FEATURES             Location/Qualifiers
     source          1..7163
                     /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             569..634
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    703..824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(714..733)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     768..787
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      939..985
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1015..1151
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1196..1267
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1493..2081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1573..1592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2581..3790
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3627..3646
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3794..4822
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4849..4897
                     /note="synthetic polyadenylation signal"
     CDS             join(5163..5231,5232..6023)
                     /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(5381..5400)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6036..6172
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6317..6388
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6393..6439)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6457..6505
                     /note="synthetic polyadenylation signal"
     enhancer        6586..6889
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6891..7093
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7044..7064
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7162..7163,1..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 agtcgatgga ttataaggac cacgatggag actacaaaga
      601 tcatgacatt gattacaagg acgacgatga caaataagga tccgcaggcc tctgctagct
      661 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
      721 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
      781 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
      841 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
      901 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
      961 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
     1021 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     1081 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     1141 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
     1201 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
     1261 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
     1321 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     1381 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     1441 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1501 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1561 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1621 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1681 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1741 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1801 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1861 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1921 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1981 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2041 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2101 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2161 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2221 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2281 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     2341 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     2401 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     2461 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     2521 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     2581 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg
     2641 ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc
     2701 ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg
     2761 acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga
     2821 aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg
     2881 attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc
     2941 gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg
     3001 gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc
     3061 aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc
     3121 agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag
     3181 gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc
     3241 gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga
     3301 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     3361 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     3421 gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg
     3481 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     3541 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     3601 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     3661 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
     3721 atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt
     3781 tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     3841 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     3901 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     3961 tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc tcccgattcc
     4021 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     4081 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     4141 cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg ggttcggccc
     4201 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     4261 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     4321 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     4381 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     4441 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     4501 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4561 gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg accaactcta
     4621 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     4681 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgccgc
     4741 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     4801 tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa taaaatcgct
     4861 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg
     4921 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     4981 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     5041 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     5101 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     5161 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5221 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5281 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5341 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5401 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5461 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5521 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5581 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5641 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5701 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     5761 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     5821 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5881 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5941 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6001 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     6061 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     6121 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt
     6181 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     6241 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     6301 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     6361 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     6421 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     6481 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     6541 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6601 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6661 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6721 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6781 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6841 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6901 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6961 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     7021 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     7081 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     7141 tcgacacacc cgccagcggc cgc
//
