LOCUS       sequence-307291-        6548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6548)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6548)
  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-m4133718
FEATURES             Location/Qualifiers
     source          1..6548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        31..334
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        336..538
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     489..509
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             976..996
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1000..1065
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1118..1239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1129..1148)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1183..1202
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1354..1400
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1430..1566
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1611..1682
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1908..2496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1988..2007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2996..4205
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4042..4061
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4209..4808
                     /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(4209..4228)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4705..4725
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4835..4883
                     /note="synthetic polyadenylation signal"
     CDS             join(5149..5217,5218..6009)
                     /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(5367..5386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6022..6158
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6310..6381
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6386..6432)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6450..6498
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
       61 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      121 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      181 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      241 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      301 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
      361 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      421 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      481 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      541 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
      601 gcggccgcca ccatgaaatt cttagtgttt cttgggatca ttacaacggt cgcagctttt
      661 catcaggaat gcagcctaca atcgtgtacc cagcaccagc cgtacgttgt ggacgatcct
      721 tgccccatcc acttctattc taagtggtac atacgggtag gcgcgaggaa gagtgcacca
      781 ctcattgagt tgtgtgtgga cgaggccggt tcaaaatccc caatccagta tatagatatt
      841 ggaaattaca ctgtctcctg tctgcccttc accatcaact gccaagaacc taagctggga
      901 agcctcgttg tgagatgctc tttttatgag gattttcttg aataccacga cgtccgtgtg
      961 gttctggact tcattgaaaa cctttacttt cagtcgatgg attataagga ccacgatgga
     1021 gactacaaag atcatgacat tgattacaag gacgacgatg acaaataagc tagcttgact
     1081 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     1141 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1201 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1261 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1321 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     1381 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     1441 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     1501 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     1561 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     1621 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     1681 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     1741 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     1801 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     1861 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1921 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1981 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2041 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2101 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2161 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2221 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2281 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2341 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2401 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2461 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2521 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2581 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2641 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2701 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     2761 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     2821 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     2881 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     2941 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct
     3001 ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg
     3061 tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
     3121 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
     3181 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
     3241 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
     3301 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
     3361 tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg
     3421 ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg
     3481 ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa
     3541 aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt
     3601 tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa
     3661 tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga
     3721 agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg
     3781 gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt
     3841 cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg
     3901 cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg
     3961 cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat
     4021 gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga
     4081 gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta
     4141 attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt
     4201 tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     4261 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     4321 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     4381 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     4441 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     4501 tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg ccgcaccggc
     4561 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     4621 gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc ggggtgcccg
     4681 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     4741 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     4801 gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg
     4861 gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc
     4921 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     4981 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     5041 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     5101 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     5161 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5221 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     5281 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     5341 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5401 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     5461 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5521 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     5581 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5641 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5701 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     5761 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5821 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5881 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5941 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6001 cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     6061 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     6121 ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg
     6181 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt
     6241 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc
     6301 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa
     6361 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa
     6421 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg
     6481 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt
     6541 acggggtc
//
