LOCUS       sequence-295911-        6691 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 6691)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6691)
  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-m4133757
FEATURES             Location/Qualifiers
     source          1..6691
                     /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..607
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    660..781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(671..690)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     725..744
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      896..942
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      972..1108
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1153..1224
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1450..2038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1530..1549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2538..3747
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3584..3603
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3751..4350
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3751..3770)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4247..4267
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4377..4425
                     /note="synthetic polyadenylation signal"
     CDS             join(4691..4759,4760..5551)
                     /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(4909..4928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5564..5700
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5845..5916
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5921..5967)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5985..6033
                     /note="synthetic polyadenylation signal"
     enhancer        6114..6417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6419..6621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6572..6592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6690..6691,1..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 cgaaagaaac cgcggcagct aagttcgaga ggcagcacat
      601 ggacagttaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
      661 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      721 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      781 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
      841 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
      901 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
      961 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1021 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1081 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1141 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1201 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1261 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     1321 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     1381 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1861 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1981 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     2281 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     2341 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     2401 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2461 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     2521 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     2581 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     2641 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     2701 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     2761 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     2821 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     2881 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     2941 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     3001 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     3061 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     3121 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     3181 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     3241 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     3301 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     3361 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     3421 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     3481 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     3541 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     3601 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     3661 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     3721 tggccgtttt tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc
     3781 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     3841 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     3901 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     3961 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4021 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag
     4081 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4141 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc
     4201 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4261 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4321 tggtgcatga cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata
     4381 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc
     4441 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     4501 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     4561 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     4621 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     4681 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4741 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4801 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4861 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4921 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4981 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     5041 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5101 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5161 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5221 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     5281 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     5341 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5401 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5461 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5521 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     5581 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     5641 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     5701 gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca
     5761 gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt
     5821 tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc
     5881 ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc
     5941 gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac
     6001 acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag
     6061 atctagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6361 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg
     6421 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     6481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     6541 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     6601 gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta
     6661 ccatccactc gacacacccg ccagcggccg c
//
