LOCUS Exported 7366 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Acute Depletion of P300-mScarlet. ACCESSION . VERSION . KEYWORDS pFlox_N-p300_Neo_FKBPF36V_mScarlet SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7366) 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 7366) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7366 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 127..149 /label=M13/pUC Forward /note="In lacZ gene" CDS 1494..2285 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1548..1567) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2158..2177 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS 2295..2351 /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 2352..2378 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2382..2408 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2418..2738 /codon_start=1 /product="F36V mutant of FK506-binding protein FKBP12" /label=FKBP-V /note="binds synthetic ligands that do not bind wild-type FKBP" /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" CDS 2769..3464 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet-I /note="matures faster than mScarlet" /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(2902..2920) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3170..3189 /label=mCherry-F /note="mCherry, forward primer" promoter complement(4869..4887) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4908..4924) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4908..4924) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4921..4943) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4932..4948 /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(4956..4986) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5001..5022 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5139..5156) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5310..5898) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5390..5409) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6069..6929) /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 6692..6711 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6930..7034) /gene="bla" /label=AmpR promoter primer_bind 7102..7120 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7158..7180) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7280..7299 /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 gacatgattg 1501 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 1561 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 1621 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 1681 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 1741 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 1801 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 1861 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 1921 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 1981 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 2041 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 2101 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 2161 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 2221 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 2281 tcttcggttc cggagctact aacttcagcc tgctgaagca ggctggagac gtggaggaga 2341 accctggacc ttacccctac gacgtgcccg actacgccgg ctacccatac gatgttccag 2401 attacgctgg aagcggagga gtgcaggtgg aaaccatctc cccaggagac gggcgcacct 2461 tccccaagcg cggccagacc tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga 2521 aagttgattc ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg 2581 tgatccgagg ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga 2641 ctatatctcc agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg 2701 ccactctcgt cttcgatgtg gagcttctaa aactggaagg tggaggcggg agtggaggtg 2761 gaggatcgat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc 2821 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 2881 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct 2941 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg 3001 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga 3061 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca 3121 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc 3181 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc 3241 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact 3301 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtggacc 3361 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct 3421 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggtggc ggtggctcgg 3481 gcggtggtgg gtcgggtggc ggcggatctg gtacccccgg gccgccttca gccaagcggc 3541 ctaaactctc atctccggcc ctctcggcgt ccgccagcga tggcacaggt tagtctcggg 3601 actcccggcc ttccatagtt cccttcgtct tttctgctcg ttgcaccttc attccagggt 3661 gcttctgtgt atctcttccc atttccccgg ccggtgttta attttaaagg tttttgaatg 3721 gactggtcga agacgtccag tagcccaacc attttctttg ccttctgata cattcattgc 3781 aacactatgt catgtcagat cgttgtgtgt tctggaatta agaggttgga agtgggagct 3841 gcggggacct ttattgttgg ctccatgcaa tttaatttgg gaaatgccgg tacaatcgtg 3901 tgtgagggct tttatgcact tttactattt ttgttactgg tgaatgaggg atttcagctc 3961 tttctcccct gttgcaatga attctttttt tctctgcttt cccaccttta atgatttttg 4021 ctcctgtgag gcagattttc tgttgaggaa tgggggcgga agacaaatct tggtgtctgc 4081 gatccaaggt tttaagtgta actaaagttc aacaaaattt cattcttagg tcccctctcc 4141 ccatccttgc ctcaagagtc agtatggacc gcagtttatt ttatatccat ttgccatgaa 4201 accaaaatgt ctcaaactgc gtgttgaggt gtagacctga ggcgtttcct ttgcagtttg 4261 tcacactctt acttagaagg gaaagaaaaa agatgaagcc tttctggaat taggtactta 4321 gaaatgggag ataataagtt gaaagactcc cagtcatgaa cttccaacta aagtgattgt 4381 acaagtttaa aaatgaacag gaggttttgt gcttgcatgt ttgttttgtt ttgctttttg 4441 gacctttgca attccagttt acaagatgaa ggaattcttt tctccagtca gcccctttgt 4501 taggagacct gaagggagac tctcccttgt ctgttctttt tcctgcttac tgttagttta 4561 gtgttattct gatgatcact gacctggtcc tctcttgctg tctgacttgt ttgagaggcc 4621 aggataatga agaagaagaa gaagaaaaag aagaagaaga agaagaatgg ctagtcatct 4681 gacagtccac tagatgttcc tccttactgc aagcatccct ttaactcaga gaagaggctc 4741 cagatctctc cttctgccaa tgagagtgta tagtggagtt aaacgtaggt accctcgagg 4801 tcgatcgacg gtatcgataa gcttgatcta gagcggccgc caccgcggtg gagctccagc 4861 ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt 4921 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 4981 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 5041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 5101 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 5161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 5221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 5281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 5341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 5401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 5461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 5521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 5581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 5761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 5881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 5941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 6541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 6841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 6901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 6961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt 7081 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc 7141 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 7201 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 7261 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 7321 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggc //