LOCUS       sequence-297374-        6326 bp    DNA     circular SYN 15-APR-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 6326)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6326)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4252778
FEATURES             Location/Qualifiers
     source          1..6326
                     /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..235
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    288..409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(299..318)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     353..372
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      524..570
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      600..736
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    781..852
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1078..1666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1158..1177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2166..3375
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3212..3231
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3379..3978
                     /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(3379..3398)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3875..3895
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4005..4053
                     /note="synthetic polyadenylation signal"
     CDS             join(4319..4387,4388..5179)
                     /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(4537..4556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5192..5328
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5480..5551
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5556..5602)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5620..5668
                     /note="synthetic polyadenylation signal"
     enhancer        5749..6052
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6054..6256
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6207..6227
                     /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 ttcagtcgta cccctatgac gtaccagatt atgcgtaagc
      241 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      301 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      361 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
      421 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
      481 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
      541 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
      601 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
      661 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
      721 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
      781 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
      841 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
      901 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
      961 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1021 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1081 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1141 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1201 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1261 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1321 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1381 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1441 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1501 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1561 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1621 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1681 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1741 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1801 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1861 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     1921 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     1981 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2041 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2101 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2161 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     2221 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     2281 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     2341 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     2401 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     2461 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     2521 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     2581 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     2641 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     2701 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     2761 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     2821 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     2881 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     2941 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     3001 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca gccggtaggt
     3061 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     3121 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     3181 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     3241 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     3301 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     3361 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     3421 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     3481 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     3541 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     3601 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     3661 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
     3721 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     3781 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
     3841 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     3901 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     3961 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     4021 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     4081 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     4141 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     4201 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     4261 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     4321 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4381 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     4441 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4501 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4561 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4621 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4681 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     4741 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4801 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4861 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     4921 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4981 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5041 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5101 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5161 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     5221 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5281 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     5341 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     5401 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     5461 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     5521 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     5581 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     5641 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     5701 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5761 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5821 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     5881 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     5941 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6001 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
     6061 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     6121 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     6181 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     6241 ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc
     6301 cactcgacac acccgccagc ggccgc
//
