LOCUS       sequence-301636-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6344)
  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-m4252829
FEATURES             Location/Qualifiers
     source          1..6344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     535..554
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             702..1301
                     /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(702..721)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1198..1218
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1328..1376
                     /note="synthetic polyadenylation signal"
     CDS             join(1642..1710,1711..2502)
                     /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(1860..1879)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2515..2651
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2803..2874
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2879..2925)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2943..2991
                     /note="synthetic polyadenylation signal"
     enhancer        3072..3375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3377..3579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3530..3550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3837..3857
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3858..3902
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    3955..4076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3966..3985)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4020..4039
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4191..4237
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4267..4403
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4448..4519
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4745..5333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4825..4844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(5833..6344,1..698)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
       61 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
      121 ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga
      181 tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
      241 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
      301 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
      361 ggtggggcca ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
      421 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
      481 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
      541 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
      601 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt
      661 ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca
      721 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
      781 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
      841 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
      901 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
      961 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     1021 aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     1081 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     1141 gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     1201 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     1261 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt
     1321 cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta
     1381 acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat
     1441 atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc
     1501 agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga
     1561 tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca
     1621 ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt
     1681 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1741 gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1801 atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg
     1861 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1921 cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat
     1981 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2041 aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2101 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2161 gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact
     2221 ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa
     2281 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2341 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2401 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2461 cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg
     2521 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     2581 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     2641 tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt
     2701 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
     2761 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
     2821 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca
     2881 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta
     2941 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg
     3001 ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca tagcccatat
     3061 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     3121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     3181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     3241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     3301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     3361 atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     3421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     3481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     3541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
     3601 agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgcc accatgtttc
     3661 atctcgttga ctttcaggtt actatagcag agatattact aattattatg aggactttta
     3721 aagtttccat ttggaatctt gattacatca taaacctcat aattaaaaat ttatctaagt
     3781 cactaactga gaataaatat tctcaattag atgaagagca accaatggag attgatgaaa
     3841 acctttactt tcagtcgaaa gaaaccgcgg cagctaagtt cgagaggcag cacatggaca
     3901 gttaagctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
     3961 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     4021 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4081 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     4141 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
     4201 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
     4261 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     4321 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     4381 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
     4441 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     4501 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     4561 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     4621 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     4681 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4741 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4801 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4861 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4921 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4981 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5041 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5101 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5161 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5221 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5281 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5341 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5401 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5461 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5521 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     5581 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     5641 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     5701 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     5761 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     5821 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     5881 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     5941 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     6001 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     6061 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     6121 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     6181 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     6241 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     6301 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggtt
//
