LOCUS       sequence-295175-        7127 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 7127)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7127)
  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-m4133561
FEATURES             Location/Qualifiers
     source          1..7127
                     /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    667..788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(678..697)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     732..751
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      903..949
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      979..1115
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1160..1231
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1457..2045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1537..1556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2545..3754
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3591..3610
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3758..4786
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4813..4861
                     /note="synthetic polyadenylation signal"
     CDS             join(5127..5195,5196..5987)
                     /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(5345..5364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6000..6136
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6281..6352
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6357..6403)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6421..6469
                     /note="synthetic polyadenylation signal"
     enhancer        6550..6853
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6855..7057
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7008..7028
                     /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 aggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag
      661 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      721 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      781 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
      841 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
      901 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
      961 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     1021 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     1081 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     1141 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     1201 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     1261 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     1321 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     1381 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1441 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1501 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1561 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1621 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1681 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1741 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1801 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1861 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1921 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1981 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2041 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2101 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2161 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2221 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2281 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     2341 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     2401 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     2461 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     2521 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     2581 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     2641 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     2701 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     2761 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     2821 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     2881 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     2941 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     3001 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     3061 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     3121 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     3181 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     3241 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     3301 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     3361 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     3421 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     3481 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     3541 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     3601 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     3661 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     3721 taaattctgg ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac
     3781 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     3841 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     3901 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     3961 tgcatccgcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     4021 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     4081 actgcccgct gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct
     4141 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     4201 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     4261 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     4321 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     4381 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     4441 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     4501 ctacttcgag cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct
     4561 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     4621 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     4681 acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga
     4741 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc
     4801 ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt
     4861 gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     4921 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     4981 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     5041 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     5101 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     5161 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5221 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     5281 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     5341 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5401 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     5461 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5521 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     5581 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5641 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     5701 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     5761 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5821 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5881 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5941 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     6001 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     6061 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     6121 agaaatttat ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     6181 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     6241 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     6301 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     6361 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     6421 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     6481 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     6541 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     6601 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     6661 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     6721 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     6781 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     6841 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     6901 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     6961 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     7021 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     7081 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
