LOCUS       Exported                6639 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252892
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6639)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6639)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6639
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             41..61
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             62..103
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    172..293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(183..202)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     237..256
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      408..454
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      484..620
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    665..736
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(962..1550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1042..1061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2050..3259
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3096..3115
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3263..4291
                     /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    4318..4366
                     /note="synthetic polyadenylation signal"
     CDS             4632..5492
                     /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(4850..4869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5505..5641
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5793..5864
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5869..5915
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5933..5981
                     /note="synthetic polyadenylation signal"
     enhancer        6062..6365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6367..6569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6520..6540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6638..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 ggggaaacct atacccaatc cacttctcgg cttagattct acctaaggat ccgcaggcct
      121 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
      181 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      241 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
      301 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
      361 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
      421 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
      481 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
      541 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
      601 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
      661 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
      721 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
      781 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
      841 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
      901 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      961 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1081 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1141 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1201 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1261 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1321 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1381 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1441 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1501 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1561 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1621 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1681 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1741 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     1801 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     1861 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     1921 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     1981 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     2041 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     2101 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     2161 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     2221 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     2281 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     2341 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     2401 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     2461 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     2521 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     2581 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     2641 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     2701 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     2761 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     2821 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     2881 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt
     2941 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     3001 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     3061 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     3121 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     3181 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     3241 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     3301 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     3361 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     3421 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     3481 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     3541 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     3601 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     3661 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     3721 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     3781 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     3841 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     3901 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     3961 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4021 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     4081 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4141 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4201 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4261 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     4321 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     4381 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     4441 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     4501 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     4561 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     4621 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4741 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4801 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4921 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5041 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5161 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     5221 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     5281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5461 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     5521 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     5581 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     5641 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     5701 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     5761 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     5821 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     5881 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     5941 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     6001 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6061 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6361 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     6601 cgatcctacc atccactcga cacacccgcc agcggccgc
//
