LOCUS       sequence-372021-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  AUTHORS   Elde NC, Child SJ, Geballe AP, Malik HS
  TITLE     Protein kinase R reveals an evolutionary model for defeating viral 
            mimicry.
  JOURNAL   Nature. 2009 Jan 22;457(7228):485-9. doi: 10.1038/nature07529. Epub 
            2008 Nov 30.
  PUBMED    19043403
REFERENCE   2  (bases 1 to 7979)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature07529"; journalName: "Nature"; date: "2009-01-22- 
            22"; volume: "457"; issue: "7228"; pages: "485-9"
COMMENT     Sequence Label: pSB819-PKR-hum
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        71..735
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    287..404
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     699..722
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(2438..2458)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2438..2456)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2477..2493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2477..2493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2490..2512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2501..2517
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2525..2531,2532..2549,2550..2555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2570..2591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2708..2725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2879..3467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2959..2978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3638..4429,4430..4498))
                     /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     4261..4280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4499..4603)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4630..5972
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5593..5640)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     6013..6031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6069..6091)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6191..6210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6234..6449
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             6450..7253
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(7384..7839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7521..7542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7733..7752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(7965..7979,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7979,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtacct
       61 tatattgaat tttcaaaaat tcttactttt tttttggatg gacgcaaaga agtttaataa
      121 tcatattaca tggcattacc accatataca tatccatata catatccata tctaatctta
      181 cttatatgtt gtggaaatgt aaagagcccc attatcttag cctaaaaaaa ccttctcttt
      241 ggaactttca gtaatacgct taactgctca ttgctatatt gaagtacgga ttagaagccg
      301 ccgagcgggt gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg
      361 tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta
      421 caatactagc ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc
      481 ttcaaatgaa cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct
      541 tatttctggg gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa
      601 tgcaaaaact gcataaccac tttaactaat actttcaaca ttttcggttt gtattacttc
      661 ttattcaaat gtaataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg
      721 tcaaggagaa aaaactatac tcgagatggc tggtgatctt tcagcaggtt tcttcatgga
      781 ggaacttaat acataccgtc agaagcaggg agtagtactt aaatatcaag aactgcctaa
      841 ttcaggacct ccacatgata ggaggtttac atttcaagtt ataatagatg gaagagaatt
      901 tccagaaggt gaaggtagat caaagaagga agcaaaaaat gccgcagcca aattagctgt
      961 tgagatactt aataaggaaa agaaggcagt tagtccttta ttattgacaa caacgaattc
     1021 ttcagaagga ttatccatgg ggaattacat aggccttatc aatagaattg cccagaagaa
     1081 aagactaact gtaaattatg aacagtgtgc atcgggggtg catgggccag aaggatttca
     1141 ttataaatgc aaaatgggac agaaagaata tagtattggt acaggttcta ctaaacagga
     1201 agcaaaacaa ttggccgcta aacttgcata tcttcagata ttatcagaag aaacctcagt
     1261 gaaatctgac tacctgtcct ctggttcttt tgctactacg tgtgagtccc aaagcaactc
     1321 tttagtgacc agcacactcg cttctgaatc atcatctgaa ggtgacttct cagcagatac
     1381 atcagagata aattctaaca gtgacagttt aaacagttct tcgttgctta tgaatggtct
     1441 cagaaataat caaaggaagg caaaaagatc tttggcaccc agatttgacc ttcctgacat
     1501 gaaagaaaca aagtatactg tggacaagag gtttggcatg gattttaaag aaatagaatt
     1561 aattggctca ggtggatttg gccaagtttt caaagcaaaa cacagaattg acggaaagac
     1621 ttacgttatt aaacgtgtta aatataataa cgagaaggcg gagcgtgaag taaaagcatt
     1681 ggcaaaactt gatcatgtaa atattgttca ctacaatggc tgttgggatg gatttgatta
     1741 tgatcctgag accagtgatg attctcttga gagcagtgat tatgatcctg agaacagcaa
     1801 aaatagttca aggtcaaaga ctaagtgcct tttcatccaa atggaattct gtgataaagg
     1861 gaccttggaa caatggattg aaaaaagaag aggcgagaaa ctagacaaag ttttggcttt
     1921 ggaactcttt gaacaaataa caaaaggggt ggattatata cattcaaaaa aattaattca
     1981 tagagatctt aagccaagta atatattctt agtagataca aaacaagtaa agattggaga
     2041 ctttggactt gtaacatctc tgaaaaatga tggaaagcga acaaggagta agggaacttt
     2101 gcgatacatg agcccagaac agatttcttc gcaagactat ggaaaggaag tggacctcta
     2161 cgctttgggg ctaattcttg ctgaacttct tcatgtatgt gacactgctt ttgaaacatc
     2221 aaagtttttc acagacctac gggatggcat catctcagat atatttgata aaaaagaaaa
     2281 aactcttcta cagaaattac tctcaaagaa acctgaggat cgacctaaca catctgaaat
     2341 actaaggacc ttgactgtgt ggaagaaaag cccagagaaa aatgaacgac acacatgtta
     2401 ggcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     2461 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2521 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2581 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4621 tgccacctga acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     4681 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     4741 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     4801 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     4861 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     4921 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     4981 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     5041 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     5101 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     5161 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     5221 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     5281 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     5341 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     5401 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     5461 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     5521 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc ttttggtttt
     5581 caaaagcgct ctgaagttcc tatactttct agagaatagg aacttcggaa taggaacttc
     5641 aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc
     5701 actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac
     5761 ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa
     5821 aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct
     5881 tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc
     5941 ttcaatgcta tcatttcctt tgatattgga tcatactaag aaaccattat tatcatgaca
     6001 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     6061 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     6121 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     6181 cttaactatg cggcatcaga gcagattgta ctgagagtgc accataccac cttttcaatt
     6241 catcattttt tttttattct tttttttgat ttcggtttcc ttgaaatttt tttgattcgg
     6301 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     6361 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     6421 caaaaacctg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     6481 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     6541 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     6601 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     6661 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag
     6721 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     6781 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     6841 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     6901 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     6961 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     7021 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     7081 acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta
     7141 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     7201 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     7261 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     7321 agttattacc ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     7381 aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     7441 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     7501 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     7561 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     7621 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     7681 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     7741 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     7801 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     7861 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     7921 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
