LOCUS       sequence-492516-        3068 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  Two Widom601 sequences, with a short linker with sequence from the 
            CDKN2B locus between them. For large scale PCR amplification of DNA 
            for nucleosome refolding..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3068)
  AUTHORS   Zhang Q, McKenzie NJ, Warneford-Thomson R, Gail EH, Flanigan SF, 
            Owen BM, Lauman R, Levina V, Garcia BA, Schittenhelm RB, Bonasio R, 
            Davidovich C
  TITLE     RNA exploits an exposed regulatory site to inhibit the enzymatic 
            activity of PRC2.
  JOURNAL   Nat Struct Mol Biol. 2019 Mar;26(3):237-247. doi: 
            10.1038/s41594-019-0197-y. Epub 2019 Mar 4.
  PUBMED    30833789
REFERENCE   2  (bases 1 to 3068)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41594-019-0197-y"; journalName: "Nat Struct Mol Biol"; 
            date: "2019-03"; volume: "26"; issue: "3"; pages: "237-247"
COMMENT     Sequence Label: pWidom601.CDKN2Bp.601
FEATURES             Location/Qualifiers
     source          1..3068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1443..1460)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1443..1459)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1452..1474)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1668..1687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1787..1809
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1847..1865)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1933..2037
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2038..2106,2107..2898)
                     /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(2256..2275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcaggcc
     1021 tctgcagtcg acgggcccgg gatccgatga tatcggatgt atatatctga cacgtgcctg
     1081 gagactaggg agtaatcccc ttggcggtta aaacgcgggg gacagcgcgt acgtgcgttt
     1141 aagcggtgct agagctgtct acgaccaatt gagcggcctc ggcaccggga ttctcgatat
     1201 ccccggggcc ggctggctcc ccactctgcc agagcgaggc ggggcagggc gccgatatcg
     1261 gatgtatata tctgacacgt gcctggagac tagggagtaa tccccttggc ggttaaaacg
     1321 cgggggacag cgcgtacgtg cgtttaagcg gtgctagagc tgtctacgac caattgagcg
     1381 gcctcggcac cgggattctc gatatcatct agatgcattc gcgaggtacc gagctcgaat
     1441 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1501 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1561 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     1621 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     1681 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1741 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1801 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     1861 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1921 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1981 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2041 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2101 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2161 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2221 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2281 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2341 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2401 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2461 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2521 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2581 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2641 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2701 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2761 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2821 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2881 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2941 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3001 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3061 ggatcttc
//
