LOCUS       Exported                6786 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252882
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6786)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6786)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6786
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    39..87
                     /note="synthetic polyadenylation signal"
     terminator      105..151
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    156..227
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      379..515
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(528..1388)
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1654..1702
                     /note="synthetic polyadenylation signal"
     CDS             complement(1729..2757)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2761..3970)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2905..2924)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4470..5058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4959..4978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5284..5355
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5400..5536
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      5566..5612
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5727..5848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5764..5783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5818..5837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5917..5958)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(5959..5979)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6237..6439)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6266..6286)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6441..6744
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gattactaga tctggctaca ctcgatcagc agttagataa taaaatcgct atccatcgaa
       61 gatggatgtg tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg
      121 ggcctctaaa cgggtcttga ggggtttttt gaattgagag gtagctgaag ctgctgttat
      181 gaccactaga gggaagaaga tacctgtgga gctaatggtc caagatgtcg ggagccgcga
      241 tcgctgccaa aaaggtaagc tgacgatagc tgacgagatt cgatgcccga cgagcaaatg
      301 agcgccacgg ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc cagagtcgag
      361 tttaaacgat tcgggatctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
      421 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
      481 agcgtcgcat caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccaa tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgcg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg
     1441 actcgattat acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc
     1501 tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga
     1561 ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc
     1621 gttctcggta caccaggcgc gcctagcgtt acacacacaa aaaaccaaca cacatccatc
     1681 ttcgatggat agcgatttta ttatctaaga ctaggcttac atactagctt attcctttgc
     1741 cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc
     1801 ggtccagacg gcggcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga
     1861 tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac
     1921 caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga gccggggcga
     1981 tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa
     2041 ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc
     2101 gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa
     2161 atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc
     2221 gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac
     2281 atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg
     2341 cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gcggcagcga tggcatccat
     2401 tgcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt
     2461 gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaact ccccaatgtc
     2521 aagcacttcc ggaatcggga gcgcggcgga tgcaaagtgc cgataaacat aacgatcttt
     2581 gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa
     2641 gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa
     2701 cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt tacccatgtt
     2761 gtctaacaaa aaagccaaaa acggccagaa tttagcggac aatttacaag tctaacactg
     2821 aaaattacat attgacccaa atgattacat ttcaaaaggt gcctaaaaaa cttcacaaaa
     2881 cacactcgcc aaccccgagc gcatagttca aaaccggagc ttcagctact taagaagata
     2941 ggtacataaa accgaccaaa gaaactgacg cctcacttat ccctcccctc accagaggtc
     3001 cggcgcctgt cgattcagga gagcctaccc taggcccgaa ccctgcgtcc tgcgacggag
     3061 aaaagcctac cgcacaccta ccggctggtg gccccaccct gcattataag ccaacagaac
     3121 gggtgacgtc acgacacgac gagggcgcgc gctcccaaag gtacgggtgc actgcccaac
     3181 ggcaccgcca taactgccgc ccccgcaaca gacgacaaac cgagttctcc agtcagtgac
     3241 aaacttcacg tcagggtccc cagatggtgc ccagcccatc tcacccgaat aagagctttc
     3301 ccgcattagc gaagggctca agaccttggg ttcttgccgc ccaccatgcc ccccaccttg
     3361 tttcaacgac ctcacagccc gcctcacaag cgtcttccat tcaagactcg ggaacagccg
     3421 ccattttgct gcgctccccc caacccccag ttcagggcaa ccttgctcgc ggacccagac
     3481 tacagccctt ggcggtctct ccacacgctt ccgtcccacc gagcggcccg gcggccacga
     3541 aagccccggc cagcccagca gcccgctact caccaagtga cgatcacagc gatccacaaa
     3601 caagaaccgc gacccaaatc ccggctgcga cggaactagc tgtgccacac ccggcgcgtc
     3661 cttatataat catcggcgtt caccgcccca cggagatccc tccgcagaat cgccgagaag
     3721 ggactacttt tcctcgcctg ttccgctctc tggaaagaaa accagtgccc tagagtcacc
     3781 caagtcccgt cctaaaatgt ccttctgctg atactggggt tctaaggccg agtcttatga
     3841 gcagcgggcc gctgtcctga gcgtccgggc ggaaggatca ggacgctcct gcgcccttcg
     3901 tctgacgtgg cagcgctcgc cgtgacgagg ggggcgcccg cgggaggcgc caaaacccgg
     3961 cgcggaggcc aggcgcgccc cgatttcgat attcctcgga tctcggatag atcgggattg
     4021 aggatgtagg ttgaacttct ctcgggtgat cccgatctcg gatattcggg attggagtaa
     4081 tacctggcga taggcgctgc caagcaatag ccaaggcgca aggccggcca agagacttgc
     4141 gatagttcgt tcgagaggta gcctgcaaga tggatcggac gaaaggtttg aagattttcc
     4201 gacgagggcc ttggagagtt cggaaattta aatgcaacta tggatgaacg aaatagacag
     4261 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4321 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4381 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4441 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4501 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4561 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4621 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4681 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4741 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4801 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4861 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4921 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4981 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5041 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5101 tggccttttg ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa
     5161 ggaccagatc gaacttgact gacgcaattc tgagcgtagt cgaaacccag acaagtcgat
     5221 ctcgccaagt agctgacttg actggttaat taagtttcga tagcccaagg taaccaatga
     5281 gattagggag ggaagaaagt ttgatgaact acttctgacc cctagtggtg tccagaaaag
     5341 accattaaag gaatgtcttg atcgaaatcg agagatcgcc tgcaggatag ctgacgacct
     5401 taaggataaa tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta
     5461 tccgttgctg aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt
     5521 ttttctgcgc caatgccggt agatagcaat ccggcccgag gggcacaaaa aacccctcaa
     5581 gacccgttta gaggccccaa ggggttatgc tacgataccg atagagatgg gccaatacca
     5641 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     5701 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     5761 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5821 gtttgtccaa actcatcaat gtatcttatc atgtctgtga gctgaaggta cgctgtatct
     5881 cagtcagtca agctagcaga ggcctgcgga tccttaggta gaatctaagc cgagaagtgg
     5941 attgggtata ggtttccccg actgaaagta aaggttttca tcaatctcca ttggttgctc
     6001 ttcatctaat tgagaatatt tattctcagt tagtgactta gataaatttt taattatgag
     6061 gtttatgatg taatcaagat tccaaatgga aactttaaaa gtcctcataa taattagtaa
     6121 tatctctgct atagtaacct gaaagtcaac gagatgaaac atggtggcgg ccgctggcgg
     6181 gtgtgtcgag tggatggtag gatcgacaaa gatctgatct gacggttcac taaaccagct
     6241 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag
     6301 ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     6361 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg
     6421 tactgccaaa accgcatcag catggtaata gcgatgacta atacgtagat gtactgccaa
     6481 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     6541 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     6601 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     6661 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     6721 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     6781 gtaatt
//
