LOCUS Exported 6943 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Acute Depletion P300-EYFP. ACCESSION . VERSION . KEYWORDS pFlox_N-p300_BSD_FKBPF36V_EYFP 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6943 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1992..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="VQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKPF KFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLK LE" CDS 2316..2342 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2343..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(4533..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(5646..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 //