LOCUS       sequence-158866-        4355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-ARS1-8xLacO (TALO8) minichromosome system for histone 
            purification in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4355)
  AUTHORS   Unnikrishnan A, Gafken PR, Tsukiyama T
  TITLE     Dynamic changes in histone acetylation regulate origins of DNA 
            replication.
  JOURNAL   Nat Struct Mol Biol. 2010 Apr;17(4):430-7. doi: 10.1038/nsmb.1780. 
            Epub 2010 Mar 14.
  PUBMED    20228802
REFERENCE   2  (bases 1 to 4355)
  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/nsmb.1780";
            journalName: "Nat Struct Mol Biol"; date: "2010-04"; volume: "17"; 
            issue: "4"; pages: "430-7"
COMMENT     Sequence Label: pUC-TALO8
FEATURES             Location/Qualifiers
     source          1..4355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    399..455
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        451..552
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             553..1227
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNS*GTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      1066..1519
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1"
     protein_bind    1883..1899
                     /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)."
     protein_bind    1910..1926
                     /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)."
     protein_bind    1937..1953
                     /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)."
     protein_bind    1964..1980
                     /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)."
     protein_bind    1991..2007
                     /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)."
     protein_bind    2018..2034
                     /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)."
     protein_bind    2045..2061
                     /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)."
     protein_bind    2072..2088
                     /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     complement(2134..2150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2134..2150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2147..2169)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2158..2174
                     /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(2182..2188,2189..2206,2207..2212))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2227..2248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2365..2382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2536..3124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2616..2635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3295..4086,4087..4155))
                     /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     3918..3937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4156..4260)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4328..4346
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atccccgggt accgagctcg aattcggtcg aaaaaagaaa aggagagggc
      481 caagagggag ggcattggtg actattgagc acgtgagtat acgtgattaa gcacacaaag
      541 gcagcttgga gtatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt
      601 tgcggcttgc agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg
      661 ggtattatat gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt
      721 tcaagtcttg taaaagcata taaaaatagt taaggcactc cgaaatactt ggttggcgtg
      781 tttcgtaatc aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc
      841 gtccaactgc atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt
      901 attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa
      961 cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat
     1021 tggaactcga tttctgactg ggttggaagg caagagagcc ccgaaagctt acattttatg
     1081 ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt
     1141 ggtgttgatg taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca
     1201 aatttcgtca aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt
     1261 ttgtgcactt gcctgcaggc cttttgaaaa gcaagcataa aagatctaaa cataaaatct
     1321 gtaaaataac aagatgtaaa gataatgcta aatcatttgg ctttttgatt gattgtacag
     1381 gaaaatatac atcgcagggg gttgactttt accatttcac cgcaatggaa tcaaacttgt
     1441 tgaagagaat gttcacaggc gcatacgcta caatgacccg attcttgcta gccttttctc
     1501 ggtcttgcaa acaaccgccg gcagcttagt atataaatac acatgtacat acctctctcc
     1561 gtatcctcgt aatcattttc ttgtatttat cgtcttttcg ctgtaaaaac tttatcacac
     1621 ttatctcaaa tacacttatt aaccgctttt actattatct tctacgctga cagtaatatc
     1681 aaacagtgac acatattaaa cacagtggtt tctttgcata aacaccatca gcctcaagtc
     1741 gtcaagtaaa gatttcgtgt tcatgcagat agataacaat ctatatgttg ataattagcg
     1801 ttgcctcatc aatgcgagat ccgtttaacc ggaccctagt gcacttaccc cacgttcggt
     1861 ccactgtgtg ccgaactagc aattgtgagc ggataacaat tactagcaat tgtgagcgga
     1921 taacaattac tagcaattgt gagcggataa caattactag caattgtgag cggataacaa
     1981 ttactagcaa ttgtgagcgg ataacaatta ctagcaattg tgagcggata acaattacta
     2041 gcaattgtga gcggataaca attactagca attgtgagcg gataacaatt actagttcct
     2101 tcactatttt aacatgtgga attcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2161 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2221 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2281 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2341 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2401 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2461 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2521 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2581 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2641 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2701 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2761 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2821 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2881 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2941 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3001 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3061 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3121 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3181 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3241 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3301 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3361 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3421 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3481 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3541 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3601 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3661 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3721 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3781 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3841 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3901 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3961 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4021 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4081 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4141 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4201 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4261 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4321 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
