LOCUS       Exported                6672 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252908
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6672)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6672
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     259..278
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             426..1454
                     /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    1481..1529
                     /note="synthetic polyadenylation signal"
     CDS             1795..2655
                     /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(2013..2032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2668..2804
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2956..3027
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      3032..3078
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3096..3144
                     /note="synthetic polyadenylation signal"
     enhancer        3225..3528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3530..3732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3683..3703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3849..3869
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3873..3938
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4007..4128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4018..4037)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4072..4091
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4243..4289
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4319..4455
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4500..4571
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4797..5385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4877..4896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5885..422
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
       61 ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt
      121 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
      181 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
      241 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
      301 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
      361 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
      421 acaacatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
      481 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
      541 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
      601 acaaagatcg ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc
      661 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
      721 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
      781 caatggatgc catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac
      841 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
      901 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
      961 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     1021 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     1081 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     1141 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     1201 gatcgccccg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     1261 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     1321 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga
     1381 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     1441 gggcaaagga ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg
     1501 aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga
     1561 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     1621 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     1681 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     1741 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     1801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     1921 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1981 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2041 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2101 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2161 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     2221 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2281 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2341 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     2401 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2461 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2521 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2581 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2641 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     2701 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     2761 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta
     2821 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     2881 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     2941 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     3001 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     3061 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     3121 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     3181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     3541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     3601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     3661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     3721 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     3781 cgacacaccc gccagcggcc gccaccatga gtgccgatgc tcagtccttt ctcaacggtt
     3841 tcgcagtgga aaacctttac tttcagtcga tggattataa ggaccacgat ggagactaca
     3901 aagatcatga cattgattac aaggacgacg atgacaaata aggatccgca ggcctctgct
     3961 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
     4021 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     4081 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     4141 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     4201 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
     4261 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
     4321 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     4381 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     4441 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
     4501 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
     4561 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
     4621 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
     4681 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
     4741 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4801 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4861 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4921 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4981 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5041 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5101 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5161 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5221 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5281 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5341 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5401 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5461 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5521 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5581 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     5641 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     5701 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     5761 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     5821 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     5881 gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc
     5941 gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga
     6001 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
     6061 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
     6121 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
     6181 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
     6241 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
     6301 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
     6361 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
     6421 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     6481 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc
     6541 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc
     6601 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     6661 gttatggcgg tg
//
