LOCUS       sequence-300745-        6854 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 6854)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6854)
  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-m4252861
FEATURES             Location/Qualifiers
     source          1..6854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        777..1986
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1823..1842
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1990..2589
                     /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(1990..2009)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2486..2506
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2616..2664
                     /note="synthetic polyadenylation signal"
     CDS             join(2930..2998,2999..3790)
                     /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(3148..3167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3803..3939
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4091..4162
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4167..4213)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4231..4279
                     /note="synthetic polyadenylation signal"
     enhancer        4360..4663
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4665..4867
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4818..4838
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5125..5145
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5179..5256
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             5284..5304
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    5753..5874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5764..5783)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5818..5837
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5989..6035
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      6065..6201
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6246..6317
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(join(6543..6854,1..277))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6623..6642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
       61 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      121 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      481 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
      541 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
      601 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
      661 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
      721 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
      781 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
      841 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
      901 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
      961 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     1021 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     1081 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     1141 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     1201 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     1261 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     1321 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     1381 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     1441 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     1501 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     1561 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     1621 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     1681 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     1741 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     1801 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     1861 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     1921 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     1981 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     2041 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     2101 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     2161 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     2221 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     2281 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     2341 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     2401 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     2461 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     2521 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     2581 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     2641 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     2701 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     2761 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     2821 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     2881 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     2941 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3001 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     3061 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     3121 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3181 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     3241 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3301 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     3361 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3421 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3481 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3541 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3601 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3661 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3721 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3781 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     3841 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     3901 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     3961 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     4021 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     4081 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     4141 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     4201 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     4261 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     4321 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     4381 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4441 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     4501 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     4561 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     4621 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     4681 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     4741 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     4801 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     4861 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     4921 cacccgccag cggccgccac catgttccac ttggtggatt ttcaggtcac gatcgcagaa
     4981 atactgctaa ttatcatgcg aaccttcaag gttagcattt ggaacctgga ctacatcatt
     5041 aatctcatca taaaaaacct tagtaagtct ttaactgaga ataaatattc ccaactggat
     5101 gaggaacagc caatggagat tgacgaaaac ctttactttc agtcgggacg gcgaatcccg
     5161 gggctgataa atccctggaa aaggagatgg aagaagaact ttattgcagt gtctgcggct
     5221 aaccggttca aaaagatcag ttcaagcggc gccctagatt acgacattcc tacgaccgcc
     5281 tccgagaatc tctatttcca gggtgaattt ggcttggcac aacatgacga ggccgtcgat
     5341 aacaagttta ataaagaaca gcagaacgct ttctatgaga tccttcactt accaaatctg
     5401 aacgaagagc aacgcaacgc tttcattcag agcctgaaag acgatccctc gcagtctgca
     5461 aatctcctgg ccgaagccaa gaagctgaat gacgcacagg ctccaaaagt tgataacaag
     5521 tttaacaagg agcagcagaa tgctttctac gagatattgc accttcctaa tctgaatgag
     5581 gaacaaagaa atgcatttat ccagtcccta aaagacgatc catcacagag tgccaacctg
     5641 cttgcggaag ccaaaaagct caatgatgcc caggccccga aagtagacgc gaaccatcaa
     5701 taagctagct tgactgactg agatacagcg taccttcagc tcacagacat gataagatac
     5761 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5821 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     5881 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     5941 aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc
     6001 ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct
     6061 accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     6121 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     6181 ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat
     6241 caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa
     6301 actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt
     6361 caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc
     6421 agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa
     6481 atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6541 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6601 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6661 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6721 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6781 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6841 aactatcgtc ttga
//
