LOCUS       sequence-295069-        6682 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 6682)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6682)
  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-m4133347
FEATURES             Location/Qualifiers
     source          1..6682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             188..208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             557..574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    651..772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(662..681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     716..735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      887..933
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      963..1099
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1144..1215
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1441..2029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1521..1540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2529..3738
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3575..3594
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3742..4341
                     /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(3742..3761)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4238..4258
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4368..4416
                     /note="synthetic polyadenylation signal"
     CDS             join(4682..4750,4751..5542)
                     /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(4900..4919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5555..5691
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5836..5907
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5912..5958)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5976..6024
                     /note="synthetic polyadenylation signal"
     enhancer        6105..6408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6410..6612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6563..6583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttt catctcgttg actttcaggt tactatagca gagatattac taattattat
       61 gaggactttt aaagtttcca tttggaatct tgattacatc ataaacctca taattaaaaa
      121 tttatctaag tcactaactg agaataaata ttctcaatta gatgaagagc aaccaatgga
      181 gattgatgaa aacctttact ttcagtcgat gagtgacaaa atcattcatc taaccgatga
      241 ctcctttgat acagacgtgt tgaaggccga tggtgcgata ctggtagatt tctgggctga
      301 atggtgtggc ccgtgcaaga tgatcgcacc cattctggac gagatcgccg acgagtatca
      361 aggaaaatta actgtcgcta agcttaatat agatcagaac ccagggacgg cacctaaata
      421 cggcatcagg ggaataccta ccctcctgct ctttaagaat ggggaagttg ccgcgacaaa
      481 agtgggagcc ctgtctaagg gccagctaaa agagttcttg gacgctaacc tggcaggttc
      541 aggctcgggg cacatgcacc atcaccatca ccatagcagc gggctcgtgc cacgcggcta
      601 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
      661 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      721 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      781 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
      841 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
      901 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
      961 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     1021 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     1081 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     1141 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     1201 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     1261 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1321 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     1381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     2281 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     2341 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     2401 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     2461 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     2521 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     2581 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     2641 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     2701 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     2761 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     2821 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     2881 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     2941 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     3001 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     3061 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     3121 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     3181 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     3241 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     3301 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     3361 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     3421 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     3481 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     3541 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     3601 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     3661 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     3721 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3781 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3841 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3901 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3961 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4021 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
     4081 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4141 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
     4201 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4261 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4321 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
     4381 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     4441 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     4501 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4561 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4621 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4681 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4741 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4801 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4861 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4921 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4981 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5041 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5101 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5161 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5221 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     5281 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5341 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5401 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5461 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5521 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5581 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5641 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcgttta
     5701 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     5761 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     5821 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     5881 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     5941 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     6001 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     6061 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     6421 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     6481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     6541 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     6601 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     6661 cgacacaccc gccagcggcc gc
//
