LOCUS       Exported                6987 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252897
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6987)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6987)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6987
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     500..519
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             667..1695
                     /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    1722..1770
                     /note="synthetic polyadenylation signal"
     CDS             2036..2896
                     /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(2254..2273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2909..3045
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3197..3268
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      3273..3319
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3337..3385
                     /note="synthetic polyadenylation signal"
     enhancer        3466..3769
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3771..3973
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3924..3944
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4042..4051
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4084..4104
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4453..4470
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4563..4684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4574..4593)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4628..4647
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4799..4845
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4875..5011
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5056..5127
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5353..5941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5433..5452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6441..663
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
       61 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
      121 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
      181 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
      241 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
      301 cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt gcggtaggct
      361 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
      421 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
      481 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
      541 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
      601 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
      661 gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
      721 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
      781 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
      841 tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat tccggaagtg
      901 cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg tgcacagggt
      961 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     1021 gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg cccattcgga
     1081 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     1141 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     1201 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     1261 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     1321 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     1381 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     1441 ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     1501 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     1561 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg
     1621 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     1681 agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     1741 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     1801 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     1861 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     1921 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     1981 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     2041 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2101 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     2161 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2221 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2281 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2341 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2401 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     2461 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2521 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2581 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     2641 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2701 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2761 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2821 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2881 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     2941 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     3001 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     3061 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     3121 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     3181 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     3241 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     3301 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     3361 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     3421 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     3481 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     3541 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     3601 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     3661 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     3721 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     3781 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     3841 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     3901 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3961 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     4021 tcgacacacc cgccagcggc cgccaccatg gctgatgcac aatcgttttt aaacgggttt
     4081 gcggaaaacc tttactttca gtcgatgagt gacaaaatca ttcatctaac cgatgactcc
     4141 tttgatacag acgtgttgaa ggccgatggt gcgatactgg tagatttctg ggctgaatgg
     4201 tgtggcccgt gcaagatgat cgcacccatt ctggacgaga tcgccgacga gtatcaagga
     4261 aaattaactg tcgctaagct taatatagat cagaacccag ggacggcacc taaatacggc
     4321 atcaggggaa tacctaccct cctgctcttt aagaatgggg aagttgccgc gacaaaagtg
     4381 ggagccctgt ctaagggcca gctaaaagag ttcttggacg ctaacctggc aggttcaggc
     4441 tcggggcaca tgcaccatca ccatcaccat agcagcgggc tcgtgccacg cggctaagga
     4501 tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc tcacagacat
     4561 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4621 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4681 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4741 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     4801 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     4861 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     4921 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     4981 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     5041 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     5101 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     5161 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     5221 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     5281 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5761 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6061 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     6181 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     6241 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     6301 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     6361 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     6421 atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     6481 cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc
     6541 gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat
     6601 cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca
     6661 gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg
     6721 tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg
     6781 tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg
     6841 agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg
     6901 aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa
     6961 ctgggggttg gggggagcgc agcaaaa
//
