LOCUS       Exported                7461 bp ds-DNA     circular SYN 26-JAN-2023
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4134063
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7461)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7461)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 26, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7461
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1456..1476
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1477..1503
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1572..1693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1583..1602)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1637..1656
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1808..1854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1884..2020
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2065..2136
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2362..2950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2442..2461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3450..4659
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4496..4515
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4663..5691
                     /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"
     polyA_signal    5718..5766
                     /note="synthetic polyadenylation signal"
     CDS             6032..6892
                     /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(6250..6269)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6905..7041
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7193..7264
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      7269..7315
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7333..7381
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagcgc cgttaagcgg
      601 acaatcaaag ggacccatca ctggttgctc cttactatac tgacgtcttt actggtcttg
      661 gtacagagta cacaatggtc gcttttcttt tttctgtacg agaatgcatt cctcccattt
      721 gcgatgggaa tcattgctat gtccgccttc gctatgatgt tcgtgaagca caaacatgcc
      781 ttcctttgcc tgtttctcct accctcactg gcaaccgtgg cctattttaa catggtgtac
      841 atgcctgcga gctgggtcat gcgaattatg acttggctgg acatggttga tacctcccta
      901 tctggcttca agttaaagga ctgtgtgatg tatgcatcag ctgtagtgct gctaatcctg
      961 atgacagctc gcacagtcta cgatgacggt gccaggagag tgtggactct gatgaatgtc
     1021 ctcaccttag tttataaagt atactatggc aacgctctcg atcaggccat tagtatgtgg
     1081 gcattgataa tctccgtgac gagtaactac agcggagtgg taaccactgt catgtttttg
     1141 gcacgcggga tagtgttcat gtgtgtcgaa tattgcccga tcttttttat cacagggaat
     1201 actctgcaat gcattatgct ggtttactgt ttcttgggct atttctgcac ctgttatttt
     1261 ggtctattct gcctgcttaa taggtacttt cgtttgacac taggagtcta tgattacttg
     1321 gtgagcacgc aggagtttcg gtatatgaac agtcagggcc tgcttccccc aaagaactct
     1381 attgacgcgt ttaaactcaa tataaagctg ttgggcgtgg gtggaaaacc ttgcatcaag
     1441 gtggccactg tgcaggaaaa cctttacttt cagtcgtacc cctatgacgt accagattat
     1501 gcgtaaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt accttcagct
     1561 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1621 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1681 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1741 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     1801 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     1861 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     1921 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     1981 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     2041 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     2101 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     2161 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     2221 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     2281 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2341 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2401 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2461 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2521 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2581 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2641 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2701 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2761 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2821 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2881 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2941 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3001 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3061 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3121 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3181 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     3241 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     3301 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     3361 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     3421 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     3481 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     3541 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     3601 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     3661 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     3721 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     3781 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     3841 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     3901 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     3961 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     4021 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     4081 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     4141 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     4201 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     4261 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     4321 gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     4381 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     4441 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     4501 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     4561 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     4621 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa aaagcctgaa
     4681 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     4741 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     4801 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     4861 cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga gttcagcgag
     4921 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     4981 accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat cgctgccgcc
     5041 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     5101 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     5161 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     5221 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     5281 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     5341 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     5401 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct ccgggcgtat
     5461 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     5521 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     5581 cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt agaagtactc
     5641 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata agctagtatg
     5701 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     5761 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     5821 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     5881 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     5941 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     6001 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     6061 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6121 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     6181 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     6241 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6301 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     6361 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6421 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     6481 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6541 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     6601 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     6661 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6721 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6781 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6841 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     6901 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     6961 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     7021 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     7081 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     7141 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     7201 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     7261 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     7321 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     7381 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     7441 tagcccatat atggagttcc g
//
