LOCUS       sequence-383841-        7744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of EGFP fused to core p300 histone acetyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7744)
  AUTHORS   Schneider MWG, Gibson BA, Otsuka S, Spicer MFD, Petrovic M, Blaukopf
            C, Langer CCH, Batty P, Nagaraju T, Doolittle LK, Rosen MK, Gerlich 
            DW
  TITLE     A mitotic chromatin phase transition prevents perforation by 
            microtubules.
  JOURNAL   Nature. 2022 Sep;609(7925):183-190. doi: 10.1038/s41586-022-05027-y.
            Epub 2022 Aug 3.
  PUBMED    35922507
REFERENCE   2  (bases 1 to 7744)
  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/s41586-022-05027-y"; journalName: "Nature"; date: 
            "2022-09"; volume: "609"; issue: "7925"; pages: "183-190"
COMMENT     Sequence Label: EGFP-p300
FEATURES             Location/Qualifiers
     source          1..7744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(160..228,229..1020)
                     /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(378..397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1191..1779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1680..1699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1933..1950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2067..2088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2103..2108,2109..2126,2127..2133)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2141..2157
                     /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     2146..2168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2165..2181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2165..2181
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2241..2260)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2307..2531
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2520..2537
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2673..3272)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2756..2776)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3253..3272
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    join(3292..3300,3301..3303,3304..3878)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3465..3484)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     3694..3711
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3948..3967)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3950..4179
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     CDS             complement(4297..4323)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4324..6174)
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      4551..4560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     6599..6618
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(6949..7218)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     CDS             complement(6952..7230)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     promoter        complement(7253..7464)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     7705..7724
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
       61 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      121 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      181 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      241 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      301 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      361 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
      421 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      481 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      541 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      601 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      661 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
      721 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      781 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      841 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
      901 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
      961 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1021 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1081 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1141 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1201 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1261 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1321 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1381 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1441 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1501 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1561 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1621 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1681 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1741 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1801 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1861 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1921 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     1981 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2041 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2101 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2161 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgacggtata
     2221 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cactcgagcc
     2281 ccagctggtt ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct
     2341 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     2401 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     2461 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     2521 agaaggcaca gtcgaggctg atcagcgagc tctagagtcg gtgggcctcg ggggcgggtg
     2581 cggggtcggc ggggccgccc cgggtggctt cggtcggagc catggggtcg tgcgctcctt
     2641 tcggtcgggc gctgcgggtc gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac
     2701 caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg
     2761 tagaagggga ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg
     2821 gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc
     2881 gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg
     2941 ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg
     3001 atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg
     3061 gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt
     3121 tcttgcagct cggtgacccg ctcgatgtgg cggtccgggt cgacggtgtg gcgcgtggcg
     3181 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc gggggacgtc gtcgcgggtg
     3241 gcgaggcgca ccgtgggctt gtactcggtc atggaaggtc gtctccttgt gggttgtggc
     3301 aagcttatca tcgtgttttt caaaggaaaa ccacgtcccc gtggttcggg gggcctagac
     3361 gtttttttaa cctcgactaa acacatgtaa agcatgtgca ccgaggcccc agatcagatc
     3421 ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg cttgaggaga
     3481 gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt tgtgccgcct
     3541 ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg ccaggtgggg
     3601 ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa gcttccagag
     3661 gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga ggggaaagac
     3721 ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca ttgccaaaag
     3781 acggcaatat ggtggaaaat aacatataga caaacgcaca ccggccttat tccaagcggc
     3841 ttcggccagt aacgttaggg gggggggagg gagagggggg gggggagagg ggcggaattg
     3901 gccgccctag atgcatgctc gacctgcagt tggacctggg agtggacacc tgtggagaga
     3961 aaggcaaagt ggatgtcatt gtcactcaag tgtatggcca gatctcaagc ctgccacacc
     4021 tcaagcttga caacaaaaag attgtctttt ctgaccagat ggacgcggcc accctcaaag
     4081 gcatcaccgc gggccaggtg aatatcaaat cctcctcgtt tttggaaact gacaatctta
     4141 gcgcagaagt catgcccgct tttgagaggg agtactcacc ccaacagctg gccctcgcag
     4201 acagcgaatt aattccagca cactggatca gttatctatg cggccgcgag ctcggatcca
     4261 ctagtaacgg ccgccagtgt gctggaattc tcaagaagcg tagtccggaa cgtcgtacgg
     4321 gtagtcctgg ctctgcgtgt gcagctccac cagcatgcac atggtggacc actgggctct
     4381 tcggagtgaa gagaactcca ggtgcttgtc ccttgccagc gtgagaaacg catcccgacc
     4441 atccatcaga tcgcagggga tgagaggatc aggatcaaca atgggaggca gggagttggc
     4501 agcagggcca gcaatgaggc ggatcacaaa gaagacctct ttatgcttct ccatggtggc
     4561 atatagtttc tgtgagaggt cgttagatac attgggcatc ccgggtttct tcttgttgcc
     4621 cctactcagg ctgctcttat ttttgctggt tttcttatta ttcttctttt tagcattttt
     4681 gctgtctccc ttggtcacat ctgtgctttc attgctggtg ttttcctctc gttttctctc
     4741 ttcttcctcc tgttccagtt ccttaatgct ttcttccaga acattgggcc agaaatcacc
     4801 ctcgaaataa ggcaattcct ttgcacttgt taatctatct tcagtagctt gtttaaaaat
     4861 atccttgtag tcatggacaa tacgctctga tacagccttg tcaagcattt ttttgtacca
     4921 ttcctgcagt cgcttgggct tgggtatctt ctggtcagga ggatggcaat ggaagatata
     4981 atcatctccc tcacttggtg gacatgccca aatatgccct gttgtgtaac ctaatttctt
     5041 gacatattct aaatatccaa ttaggatttc atgatagact gcagtcctca agcatttagg
     5101 acggaagaaa tgaacactat cgaggtaaga tatgtatact ctcctctggt tgggtggagg
     5161 gcagtcagag ccatactctt gaacatgcat gccaaagaag cacaggtcaa caccatcaat
     5221 ttcttcaaag gcaaagaggg ctttggttcg gtatggaaag gattctgcca tctctccact
     5281 gtccacaaac cttgctttca tgcctggttt tacttccacg gttttgtcag aagcatgaac
     5341 tactctaaca gtgacctctc ctgactcagg gtgattctgt cgcctcagaa agtcattcac
     5401 acgattctct agaaaggtgc caagtctggt agatggcaac cttttagcag aaaacttatt
     5461 ttctttccta gttcgtgcac ttttctttaa acagccatca cagacgaatc cagcaggcca
     5521 gatgatctca tggtgaagga cacagatctg atgcatcttt cttccgcact ctgtacattc
     5581 aacaaacagt tcaggatcca gtgtgtcatt ttttctcttg gaaaattgtt ctttatttat
     5641 tgtagtttga ggctgggaag ggtcatcccc caaagaaacg ctctcccctt ggatctcatt
     5701 gaaacacttc tcacagaaat gatacctgtt ctggtaactg taataagtgg catcacgagg
     5761 tattgtgcac aactgtttgc cgtagcaaca cagtgtctgt ggagagaact ccaactttct
     5821 gccacaacag tatccaaggc tttgcatcac tgggtcaatt tcttgttcaa agacctcaga
     5881 gagcttggag cagtatttgt atacccgtga tgttttccgg ttatataacc aggcattatt
     5941 gaacataagc caaatatcat cgacatactg ccagggctcc tgatactgtc cagtgtctaa
     6001 cttcctctta atggtagaaa gatccatggg gctcttcaca atatcaaagt aatcagggat
     6061 tcctaaaagc tgagggtcca caggttgacg aaagggaagg gattctggat cctgacggta
     6121 aagtgcctcc aaagttggca tcagtgcctg tcgtagttct tctggtttga aaatggcgcg
     6181 cccgctagcc ttggatccgg caattggctt gtacagctcg tccatgccga gagtgatccc
     6241 ggctgcggtc acgaactcct taaggaccat gtgatctcgc ttctcgttgg ggtctttgct
     6301 cagggcggac tgggtgctca ggtagtggtt gtcaggcagc agcacgggtc cgtcaccgat
     6361 tggggtgttc tgctggtagt ggtcggcgag ctgcacgctt ccgtcctcga tgttgtgacg
     6421 gatcttgaag ttcaccttga ttccgttctt ctgcttgtca gccatgatat agacgttgtg
     6481 gctgttgtag ttgtactcca gcttgtgacc caggatgtta ccgtcctcct tgaagtcgat
     6541 gcccttcagc tcgatgcggt tcaccagggt gtctccctcg aacttcacct cagcgcgagt
     6601 cttgtagttg ccgtcgtcct tgaagaagat ggtacgctcc tggacgtagc cttcgggcat
     6661 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tatcggctga agcactgcac
     6721 gccgtaggtc agggtggtca cgagggtagg ccagggcacg ggcagcttgc cggtggtgca
     6781 gatgaacttc agggtcagct tgccgtaggt agcatcgccc tctccctcac cactcacact
     6841 gaacttgtga ccgtttacgt caccgtccag ttcgaccagg attggcacca ctccggtgaa
     6901 cagctcctct cccttgctca cggatcctcc acctccacct ccgccggcct gctttgagat
     6961 tcgtcggaac acatgataat agaggtccca ggcttgggtg aggtccttga cattccctga
     7021 tttcatgtac ttcctgcacc actcttgggc ctccattaaa tctcgaccat aggcctgatt
     7081 aaaggatgtt tccttcagag tctgggggcc ccgttccatc atagcatgca agggctccag
     7141 cacctcaaac atgcctttca cgttcctttc cccaaagtac aaacgagatg cctcttccag
     7201 gccttcatgc cacatctcat gccagaggat catggtggct ctagaaccgg tcctgtgttc
     7261 tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt atatacggtt
     7321 ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc cagtttaccc
     7381 cgcgccacct tctctaggca ccggatcaat tgccgacccc tccccccaac ttctcgggga
     7441 ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gccaattccc actcctttca
     7501 agacctagct agcactagtc aataatcaat gtcaacgcgt atatctggcc cgtacatcgc
     7561 gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt
     7621 gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag
     7681 ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     7741 gggg
//
