LOCUS       sequence-316816-        6367 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 6367)
  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 6367)
  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: pSB305-K3L-vac
FEATURES             Location/Qualifiers
     source          1..6367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        183..847
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    399..516
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     811..834
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(1156..1175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1157..1175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1182..1199)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1182..1198)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1191..1213)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1339..1794
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1426..1445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1636..1657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2095..2502
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2503..3597
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGSGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4091..4110)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4210..4232
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4270..4288)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4356..4460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4461..4529,4530..5321)
                     /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(4679..4698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5492..6080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5981..6000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6234..6251
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggtac cttatattga
      181 attttcaaaa attcttactt tttttttgga tggacgcaaa gaagtttaat aatcatatta
      241 catggcatta ccaccatata catatccata tacatatcca tatctaatct tacttatatg
      301 ttgtggaaat gtaaagagcc ccattatctt agcctaaaaa aaccttctct ttggaacttt
      361 cagtaatacg cttaactgct cattgctata ttgaagtacg gattagaagc cgccgagcgg
      421 gtgacagccc tccgaaggaa gactctcctc cgtgcgtcct cgtcttcacc ggtcgcgttc
      481 ctgaaacgca gatgtgcctc gcgccgcact gctccgaaca ataaagattc tacaatacta
      541 gcttttatgg ttatgaagag gaaaaattgg cagtaacctg gccccacaaa ccttcaaatg
      601 aacgaatcaa attaacaacc ataggatgat aatgcgatta gttttttagc cttatttctg
      661 gggtaattaa tcagcgaagc gatgattttt gatctattaa cagatatata aatgcaaaaa
      721 ctgcataacc actttaacta atactttcaa cattttcggt ttgtattact tcttattcaa
      781 atgtaataaa agtatcaaca aaaaattgtt aatatacctc tatactttaa cgtcaaggag
      841 aaaaaactat actcgagatg cttgcatttt gttattcgtt gcccaatgcg ggcgatgtaa
      901 taaagggcag agtatacgag aaggattatg ctctatacat ttatcttttt gactatcctc
      961 actctgaagc tatcttggca gagagtgtta agatgcatat ggatagatat gttgaatata
     1021 gggataaact ggtagggaaa actgtaaaag ttaaagtgat tagagttgat tatacaaaag
     1081 gatatataga tgtcaattac aaaaggatgt gtagacatca ataagcggcc gccaccgcgg
     1141 tggagctcca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     1201 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1261 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1321 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1741 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     1801 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatcga ccggtcgagg
     1861 agaacttcta gtatatctac atacctaata ttattgcctt attaaaaatg gaatcccaac
     1921 aattacatca aaatccacat tctcttcaaa atcaattgtc ctgtacttcc ttgttcatgt
     1981 gtgttcaaaa acgttatatt tataggataa ttatactcta tttctcaaca agtaattggt
     2041 tgtttggccg agcggtctaa ggcgcctgat tcaagaaata tcttgaccgc agttaactgt
     2101 gggaatactc aggtatcgta agatgcaaga gttcgaatct cttagcaacc attatttttt
     2161 tcctcaacat aacgagaaca cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg
     2221 aaattttttg ttaatttcag aggtcgcctg acgcatatac ctttttcaac tgaaaaattg
     2281 ggagaaaaag gaaaggtgag agcgccggaa ccggcttttc atatagaata gagaagcgtt
     2341 catgactaaa tgcttgcatc acaatacttg aagttgacaa tattatttaa ggacctattg
     2401 ttttttccaa taggtggtta gcaatcgtct tactttctaa cttttcttac cttttacatt
     2461 tcagcaatat atatatatat atttcaagga tataccattc taatgtctgc ccctaagaag
     2521 atcgtcgttt tgccaggtga ccacgttggt caagaaatca cagccgaagc cattaaggtt
     2581 cttaaagcta tttctgatgt tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt
     2641 ggtggtgctg ctatcgatgc tacaggtgtt ccacttccag atgaggcgct ggaagcctcc
     2701 aagaaggctg atgccgtttt gttaggtgct gtgggtggtc ctaaatgggg ttccggtagt
     2761 gttagacctg aacaaggttt actaaaaatc cgtaaagaac ttcaattgta cgccaactta
     2821 agaccatgta actttgcatc cgactctctt ttagacttat ctccaatcaa gccacaattt
     2881 gctaaaggta ctgacttcgt tgttgtcaga gaattagtgg gaggtattta ctttggtaag
     2941 agaaaggaag acgatggtga tggtgtcgct tgggatagtg aacaatacac cgttccagaa
     3001 gtgcaaagaa tcacaagaat ggccgctttc atggccctac aacatgagcc accattgcct
     3061 atttggtcct tggataaagc taatgttttg gcctcttcaa gattatggag aaaaactgtg
     3121 gaggaaacca tcaagaacga attccctaca ttgaaggttc aacatcaatt gattgattct
     3181 gccgccatga tcctagttaa gaacccaacc cacctaaatg gtattataat caccagcaac
     3241 atgtttggtg atatcatctc cgatgaagcc tccgttatcc caggttcctt gggtttgttg
     3301 ccatctgcgt ccttggcctc tttgccagac aagaacaccg catttggttt gtacgaacca
     3361 tgccacggtt ctgctccaga tttgccaaag aataaggtca accctatcgc cactatcttg
     3421 tctgctgcaa tgatgttgaa attgtcattg aacttgcctg aagaaggtaa ggccattgaa
     3481 gatgcagtta aaaaggtttt ggatgcaggt atcagaactg gtgatttagg tggttccaac
     3541 agtaccaccg aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa
     3601 gattctcttt ttttatgata tttgtacata aactttataa atgaaattca taatagaaac
     3661 gacacgaaat tacaaaatgg aatatgttca tagggtagac gaaactatat acgcaatcta
     3721 catacattta tcaagaagga gaaaaaggag gatgtaaagg aatacaggta agcaaattga
     3781 tactaatggc tcaacgtgat aaggaaaaag aattgcactt taacattaat attgacaagg
     3841 aggagggcac cacacaaaaa gttaggtgta acagaaaatc atgaaactat gattcctaat
     3901 ttatatattg gaggattttc tctaaaaaaa aaaaaataca acaaataaaa aacactcaat
     3961 gacctgacca tttgatggag tttaagtcaa taccttcttg aaccatttcc cataatggtg
     4021 aaagttccct caagaatttt actctgtcag aaacggcctt aacgacgtag tcgatatggt
     4081 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4141 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4201 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4261 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4321 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4681 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4981 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5161 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5341 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5401 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6121 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6181 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6241 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6301 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6361 acgcaat
//
