LOCUS       sequence-307326-        6317 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 6317)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6317)
  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-m4252851
FEATURES             Location/Qualifiers
     source          1..6317
                     /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             209..226
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    279..400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(290..309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     344..363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      515..561
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      591..727
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    772..843
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1069..1657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1149..1168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2157..3366
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3203..3222
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3370..3969
                     /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(3370..3389)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3866..3886
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3996..4044
                     /note="synthetic polyadenylation signal"
     CDS             join(4310..4378,4379..5170)
                     /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(4528..4547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5183..5319
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5471..5542
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5547..5593)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5611..5659
                     /note="synthetic polyadenylation signal"
     enhancer        5740..6043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6045..6247
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6198..6218
                     /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 ttcagtcgca ccatcaccat caccattaag ctagcttgac
      241 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      301 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      361 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      421 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      481 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      541 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      601 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      661 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      721 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      781 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      841 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      901 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      961 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1021 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1081 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1141 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1201 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1261 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1321 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1381 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1441 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1501 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1561 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1621 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1681 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1741 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1801 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1861 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1921 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1981 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2041 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2101 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2161 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2221 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2281 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2341 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2401 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2461 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2521 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2581 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2641 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2701 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     2761 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     2821 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     2881 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     2941 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     3001 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     3061 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3121 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3181 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3241 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3301 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3361 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3421 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3481 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3541 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3601 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3661 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3721 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3781 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     3841 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3901 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     3961 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     4021 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     4081 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4141 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4201 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4261 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4321 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4381 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4441 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4501 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4561 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4621 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4681 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4741 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4801 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4861 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4921 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4981 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5041 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5101 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5161 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5221 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5281 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5341 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5401 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     5461 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     5521 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     5581 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     5641 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5701 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5761 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5821 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5881 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5941 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6001 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     6061 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6121 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6181 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6241 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6301 cacccgccag cggccgc
//
