LOCUS       sequence-297299-        6771 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 6771)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6771)
  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-m4252902
FEATURES             Location/Qualifiers
     source          1..6771
                     /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    304..425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(315..334)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     369..388
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      540..586
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      616..752
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    797..868
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1094..1682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1174..1193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2182..3391
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3228..3247
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3395..4423
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4450..4498
                     /note="synthetic polyadenylation signal"
     CDS             join(4764..4832,4833..5624)
                     /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(4982..5001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5637..5773
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5925..5996
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6001..6047)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6065..6113
                     /note="synthetic polyadenylation signal"
     enhancer        6194..6497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6499..6701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6652..6672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc cacttggtgg attttcaggt cacgatcgca gaaatactgc taattatcat
       61 gcgaaccttc aaggttagca tttggaacct ggactacatc attaatctca tcataaaaaa
      121 ccttagtaag tctttaactg agaataaata ttcccaactg gatgaggaac agccaatgga
      181 gattgacgaa aacctttact ttcagtcgta cccctatgac gtaccagatt atgcgtaagg
      241 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
      301 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      361 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      421 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      481 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
      541 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
      601 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
      661 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
      721 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
      781 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
      841 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
      901 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
      961 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     1021 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1501 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1561 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1621 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1681 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1741 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1801 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1861 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     1921 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     1981 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     2041 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     2101 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     2161 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     2221 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     2281 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     2341 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     2401 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     2461 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     2521 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     2581 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     2641 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     2701 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     2761 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     2821 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     2881 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     2941 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     3001 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     3061 ccaccagccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     3121 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     3181 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     3241 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     3301 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     3361 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     3421 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     3481 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     3541 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     3601 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     3661 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     3721 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     3781 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     3841 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     3901 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     3961 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     4021 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     4081 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     4141 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     4201 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     4261 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     4321 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     4381 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     4441 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     4501 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     4561 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     4621 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     4681 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     4741 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     4801 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4861 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     4921 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     4981 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5041 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     5101 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5161 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     5221 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5281 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     5341 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     5401 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5461 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5521 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5581 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     5641 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     5701 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     5761 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     5821 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     5881 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     5941 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
     6001 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
     6061 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
     6121 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
     6181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     6301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     6361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     6421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     6481 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     6541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     6601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     6661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     6721 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
