LOCUS       Exported                6576 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252982
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6576)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6576
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    109..230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(120..139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     174..193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      345..391
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      421..557
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    602..673
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(899..1487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(979..998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1987..3196
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3033..3052
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3200..4228
                     /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    4255..4303
                     /note="synthetic polyadenylation signal"
     CDS             4569..5429
                     /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(4787..4806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5442..5578
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5730..5801
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      5806..5852
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5870..5918
                     /note="synthetic polyadenylation signal"
     enhancer        5999..6302
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6304..6506
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6457..6477
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6575..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg taaggatccg caggcctctg
       61 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
      121 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      181 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      241 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
      301 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
      361 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
      421 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
      481 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
      541 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
      601 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
      661 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
      721 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
      781 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
      841 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      901 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      961 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1021 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1081 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1141 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1201 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1261 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1321 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1381 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1441 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1501 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1561 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1621 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1681 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     1741 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     1801 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     1861 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     1921 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     1981 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     2041 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     2101 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     2161 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     2221 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     2281 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     2341 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     2401 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     2461 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     2521 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     2581 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     2641 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     2701 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     2761 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     2821 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg
     2881 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     2941 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     3001 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     3061 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     3121 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     3181 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     3241 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     3301 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     3361 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     3421 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     3481 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     3541 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     3601 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     3661 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     3721 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     3781 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     3841 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     3901 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     3961 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     4021 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     4081 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     4141 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     4201 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     4261 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     4321 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     4381 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     4441 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     4501 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     4561 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4621 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4681 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4741 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4801 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4861 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4921 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4981 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5041 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5101 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     5161 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     5221 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5281 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5341 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5401 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     5461 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     5521 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     5581 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     5641 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     5701 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     5761 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     5821 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     5881 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     5941 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     6001 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     6061 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     6121 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     6181 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     6241 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     6301 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     6361 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     6421 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     6481 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     6541 tcctaccatc cactcgacac acccgccagc ggccgc
//
