LOCUS       Exported                6170 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252796
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6170)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6170)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6170
                     /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..79
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    132..253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(143..162)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     197..216
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      368..414
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      444..580
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    625..696
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(922..1510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1002..1021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2010..3219
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3056..3075
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3223..3822
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3223..3242)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3719..3739
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3849..3897
                     /note="synthetic polyadenylation signal"
     CDS             4163..5023
                     /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(4381..4400)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5036..5172
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5324..5395
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5400..5446
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5464..5512
                     /note="synthetic polyadenylation signal"
     enhancer        5593..5896
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5898..6100
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6051..6071
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6169..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gcaccatcac catcaccatt aagctagctt gactgactga gatacagcgt accttcagct
      121 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      181 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      241 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
      301 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
      361 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
      421 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
      481 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
      541 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
      601 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
      661 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
      721 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
      781 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
      841 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1741 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     1801 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     1861 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     1921 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     1981 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     2041 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     2101 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     2161 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     2221 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     2281 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     2341 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     2401 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     2461 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     2521 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     2581 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     2641 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     2701 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     2761 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     2821 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     2881 gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     2941 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     3001 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     3061 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     3121 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     3181 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc
     3241 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3301 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3361 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3421 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3481 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3541 caacagatgg agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3601 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3661 ggagtggagg ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3721 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3781 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag
     3841 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     3901 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     3961 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     4021 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     4081 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     4141 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4201 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4261 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     4321 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     4381 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4441 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     4501 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4561 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     4621 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4681 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     4741 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     4801 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4861 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4921 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4981 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     5041 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     5101 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     5161 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     5221 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     5281 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     5341 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     5401 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     5461 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     5521 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     5581 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5641 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5701 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5761 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5821 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5881 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     5941 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     6001 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     6061 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     6121 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
