LOCUS       sequence-323513-        6943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acute Depletion P300-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6943)
  AUTHORS   Zhu Y, Denholtz M, Lu H, Murre C
  TITLE     Calcium signaling instructs NIPBL recruitment at active enhancers 
            and promoters via distinct mechanisms to reconstruct genome 
            compartmentalization.
  JOURNAL   Genes Dev. 2021 Jan 1;35(1-2):65-81. doi: 10.1101/gad.343475.120. 
            Epub 2020 Dec 17.
  PUBMED    33334824
REFERENCE   2  (bases 1 to 6943)
  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"; journalName: "Genes 
            Dev."; date: "2021-01-1"; volume: "35(1-2):65-81. doi"; pages: " 
            10.1101/gad.343475.120. Epub 2020 Dec 17"
COMMENT     Sequence Label: pFlox_N-p300_BSD_FKBPF36V_EYFP
FEATURES             Location/Qualifiers
     source          1..6943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1494..1889
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1899..1955
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1956..1979
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1989..2309
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2316..2342
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2343..2345,2346..3056)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2385..2406)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2646..2665)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2993..3014
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4446..4464)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4485..4501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4485..4501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4498..4520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4509..4525
                     /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(4533..4539,4540..4557,4558..4563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4578..4599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4716..4733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4887..5475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4967..4986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5646..6437,6438..6506))
                     /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     6269..6288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6507..6611)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6679..6697
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6735..6757)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6857..6876
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccgcggc ggccgcacta gtggccgcgg
      181 cggccgcact agtggccgcg gcggccgcag cttgcctgca ggtcgacggc agaggcaaga
      241 aacaacagcc gccatcttgt ttgtgtgcta gcctgtgggg ggggtggggg gaggaggaga
      301 gagagcggag agggaccggg agagagagag tggggcaagc gggacgcccg gctgagtgcg
      361 aactgcgcga gcagggagtg aggagggtac ctggctggag agcagcggcc gggaccgaga
      421 cagtggttgg gggccccggg gcgcaaaggc tgaggcgacc ctcagtcccc tccttgcccg
      481 cacacacccc caccgcagcc cgggccggcg tcgacgccta agggggggaa accattacat
      541 aagcccgcgc cccgcggcgc ctctgcccgc cgtccgaggc ggcccccgag ccgagccccc
      601 ggcaggcacc tggctgccgg gggggtgagg gccagggagg cggccggact gatttctcct
      661 ccccgggctt gcgagtggag gcggagaagc gcggcgcgcg gagaggccgc gtcggtgggc
      721 tcggcgtcag cgcgggcttg cgggcggacg gccctgcttg gcccggcccg gcccccctgg
      781 cccttcgggc cggcgagctc cacgctcggc gtcgtcgctg ctgcgcctct gcttgtccgc
      841 tgcgcgatgt gaggcggccc cggcgccagt ctggctctcg gctcggctga gctcagtgtg
      901 gccattaggg gccggtgcgg cagcggcggc gcggagcgca gcggcaggag gaggaggagg
      961 gcggctctca gggtgggggc gcaggcccgg gagggggcac cgggaggagg tgagtgtctc
     1021 tggtcgcctc ctcctctccc cctttttcgc ccccgcctcc ttgtggcgat gagaaggagg
     1081 aggacagcgc cgaggaggaa gaagctgatg gcggcggcgg agctccgaga gacctcggct
     1141 gggcaggggc tcgccgtggc gggctgggga ctgcgtctgt agagctgtga gttcccgaga
     1201 attcgctgca gcggacgtgc tcgtgggatc agtgttgctg taccctcctc ccggcctggt
     1261 ttaggcccgg cccctcgcac ttgcccctac catttctgtt gagtccgcat ccctctccgg
     1321 ccaccgcaac caggcgaaga gaaaaaggaa cttcccccca cccctgttgg gtggcggcgg
     1381 agctccggag cccaccctta ggagcggggc ggggaccccg tttcggagaa gagatttcct
     1441 caggattctc gtttttttcc tcggttgtat ctccgaaaga attaaaagtc gacatggcca
     1501 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     1561 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     1621 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     1681 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     1741 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     1801 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     1861 tgctgccctc tggttatgtg tgggagggcg gttccggagc cacgaacttc tctctgttaa
     1921 agcaagcagg agacgtggaa gaaaaccccg gtcccgatta caaggatgac gatgacaagg
     1981 gttccggcgg cgtgcaggtg gagactatct ccccaggaga cgggcgcacc ttccccaagc
     2041 gcggccagac ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaagttgatt
     2101 cctcccggga cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag
     2161 gctgggaaga aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc
     2221 cagattatgc ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg
     2281 tcttcgatgt ggagcttcta aaactggaag tcgattatcc gtacgacgta ccagactacg
     2341 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2401 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2461 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2521 tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     2581 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2641 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2701 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2761 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2821 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2881 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2941 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3001 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtg
     3061 gaggtggatc tggtggcggc ggttcaggta cccccgggcc gccttcagcc aagcggccta
     3121 aactctcatc tccggccctc tcggcgtccg ccagcgatgg cacaggttag tctcgggact
     3181 cccggccttc catagttccc ttcgtctttt ctgctcgttg caccttcatt ccagggtgct
     3241 tctgtgtatc tcttcccatt tccccggccg gtgtttaatt ttaaaggttt ttgaatggac
     3301 tggtcgaaga cgtccagtag cccaaccatt ttctttgcct tctgatacat tcattgcaac
     3361 actatgtcat gtcagatcgt tgtgtgttct ggaattaaga ggttggaagt gggagctgcg
     3421 gggaccttta ttgttggctc catgcaattt aatttgggaa atgccggtac aatcgtgtgt
     3481 gagggctttt atgcactttt actatttttg ttactggtga atgagggatt tcagctcttt
     3541 ctcccctgtt gcaatgaatt ctttttttct ctgctttccc acctttaatg atttttgctc
     3601 ctgtgaggca gattttctgt tgaggaatgg gggcggaaga caaatcttgg tgtctgcgat
     3661 ccaaggtttt aagtgtaact aaagttcaac aaaatttcat tcttaggtcc cctctcccca
     3721 tccttgcctc aagagtcagt atggaccgca gtttatttta tatccatttg ccatgaaacc
     3781 aaaatgtctc aaactgcgtg ttgaggtgta gacctgaggc gtttcctttg cagtttgtca
     3841 cactcttact tagaagggaa agaaaaaaga tgaagccttt ctggaattag gtacttagaa
     3901 atgggagata ataagttgaa agactcccag tcatgaactt ccaactaaag tgattgtaca
     3961 agtttaaaaa tgaacaggag gttttgtgct tgcatgtttg ttttgttttg ctttttggac
     4021 ctttgcaatt ccagtttaca agatgaagga attcttttct ccagtcagcc cctttgttag
     4081 gagacctgaa gggagactct cccttgtctg ttctttttcc tgcttactgt tagtttagtg
     4141 ttattctgat gatcactgac ctggtcctct cttgctgtct gacttgtttg agaggccagg
     4201 ataatgaaga agaagaagaa gaaaaagaag aagaagaaga agaatggcta gtcatctgac
     4261 agtccactag atgttcctcc ttactgcaag catcccttta actcagagaa gaggctccag
     4321 atctctcctt ctgccaatga gagtgtatag tggagttaaa cgtaggtacc ctcgaggtcg
     4381 atcgacggta tcgataagct tgatctagag cggccgccac cgcggtggag ctccagcttt
     4441 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     4501 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4561 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4621 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4681 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4741 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4801 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4861 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4921 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4981 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5041 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5101 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5161 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5221 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5281 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5341 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5401 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5461 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5521 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5581 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5641 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5701 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5761 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5821 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5881 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5941 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6001 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6061 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6121 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6181 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6241 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6301 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6361 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6421 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6481 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6541 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6601 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6661 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     6721 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     6781 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     6841 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     6901 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggc
//
