LOCUS       sequence-491704-        6924 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Expresses histone H4 fused with photoactivatable-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  AUTHORS   Hihara S, Pack CG, Kaizu K, Tani T, Hanafusa T, Nozaki T, Takemoto 
            S, Yoshimi T, Yokota H, Imamoto N, Sako Y, Kinjo M, Takahashi K, 
            Nagai T, Maeshima K
  TITLE     Local nucleosome dynamics facilitate chromatin accessibility in 
            living mammalian cells.
  JOURNAL   Cell Rep. 2012 Dec 27;2(6):1645-56. doi: 
            10.1016/j.celrep.2012.11.008. Epub 2012 Dec 13.
  PUBMED    23246002
REFERENCE   2  (bases 1 to 6924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2012.11.008"; journalName: "Cell Rep"; date: 
            "2012-12-27- 27"; volume: "2"; issue: "6"; pages: "1645-56"
COMMENT     Sequence Label: pEF1-PA-GFP-H4-FRT
FEATURES             Location/Qualifiers
     source          1..6924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..76
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     310..327
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    444..465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(480..485,486..503,504..510)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    518..534
                     /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     523..545
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     542..558
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     542..558
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    607..728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(618..637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     672..691
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(858..1880)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(858..1880)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(1888..1935)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     2067..2086
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2219..2443
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2432..2449
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2487..2528)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(join(2949..3659,3660..3662,3663..3665))
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2991..3012)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3340..3359
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3599..3620
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3662..3671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3760..3779)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3761..3779)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3796..4974)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3805..4743
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3828..3848)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     5262..5281
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5356..5460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5461..5529,5530..6321)
                     /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(5679..5698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6492..6924,1..156)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
       61 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      121 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      181 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      241 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      301 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
      361 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
      421 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
      481 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      541 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag
      601 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      661 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      721 aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
      781 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
      841 aaatctcgta gtacgtgcta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca
      901 ctatcggcga gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt
      961 tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc
     1021 caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg
     1081 cggagcatat acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag
     1141 tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc
     1201 tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca
     1261 ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg
     1321 cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg
     1381 tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca
     1441 cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg
     1501 ctaagatcgg ccgcagcgat cgcatccatg gcctccgcga ccggctgcag aacagcgggc
     1561 agttcggttt caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag
     1621 gtcaggctct cgctgaattc cccaatgtca agcacttccg gaatcgggag cgcggccgat
     1681 gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc
     1741 aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag
     1801 agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc
     1861 gcggtgagtt caggcttttt ggccaaggaa gttcctatac tttctagaga ataggaactt
     1921 cggaatagga acttctaggt acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1981 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2041 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2101 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2161 ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc ctcagaagcc
     2221 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
     2281 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     2341 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     2401 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg
     2461 gtttaaactc attactaacc ggtacgcgta gaatcgagac cgaggagagg gttagggata
     2521 ggcttacctt cgaaccgcgg gccctctaga ctcgagcggc cgccactgtg ctggatgatc
     2581 ccgggtcagc cgccaaagcc atacagagtg cgcccctgac gtttcagggc atatactaca
     2641 tccatggctg tgacagtttt gcgcttggcg tgctccgtat aggtgacggc gtctcgaata
     2701 acgttctcta agaaaacctt aagcacacct cgagtctcct catagataag accggaaatg
     2761 cgcttgacgc caccgcgccg agccaaacgg cgaatagccg gttttgtaat gccctggatg
     2821 ttatcccgga gcaccttacg atggcgctta gcaccaccct tccccaagcc ttttccgcct
     2881 ttgccgcgcc cagacatggt accgtcgact gcagaattcg aagcttgagc tcgagatctg
     2941 agtccggact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     3001 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtggctc
     3061 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     3121 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttggccttg
     3181 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     3241 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     3301 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     3361 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     3421 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtagct gaaggtggtc
     3481 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     3541 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     3601 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     3661 accatggtgg cgaccggtag cgctagatct gcagaattcc accacactgg actagtggat
     3721 ccgagctcgg taccaagctt acctagccag cttgggtctc cctatagtga gtcgtattag
     3781 taccaagcta attcctcacg acacctgaaa tggaagaaaa aaactttgaa ccactgtctg
     3841 aggcttgaga atgaaccaag atccaaactc aaaaagggca aattccaagg agaattacat
     3901 caagtgccaa gctggcctaa cttcagtctc cacccactca gtgtggggaa actccatcgc
     3961 ataaaacccc tccccccaac ctaaagacga cgtactccaa aagctcgaga actaatcgag
     4021 gtgcctggac ggcgcccggt actccgtgga gtcacatgaa gcgacggctg aggacggaaa
     4081 ggcccttttc ctttgtgtgg gtgactcacc cgcccgctct cccgagcgcc gcgtcctcca
     4141 ttttgagctc cctgcagcag ggccgggaag cggccatctt tccgctcacg caactggtgc
     4201 cgaccgggcc agccttgccg cccagggcgg ggcgatacac ggcggcgcga ggccaggcac
     4261 cagagcaggc cggccagctt gagactaccc ccgtccgatt ctcggtggcc gcgctcgcag
     4321 gccccgcctc gccgaacatg tgcgctggga cgcacgggcc ccgtcgccgc ccgcggcccc
     4381 aaaaaccgaa ataccagtgt gcagatcttg gcccgcattt acaagactat cttgccagaa
     4441 aaaaagcgtc gcagcaggtc atcaaaaatt ttaaatggct agagacttat cgaaagcagc
     4501 gagacaggcg cgaaggtgcc accagattcg cacgcggcgg ccccagcgcc caggccaggc
     4561 ctcaactcaa gcacgaggcg aaggggctcc ttaagcgcaa ggcctcgaac tctcccaccc
     4621 acttccaacc cgaagctcgg gatcaagaat cacgtactgc agccaggtgg aagtaattca
     4681 aggcacgcaa gggccataac ccgtaaagag gccaggcccg cgggaaccac acacggcact
     4741 tacctgtgtt ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga ctactgcact
     4801 tatatacggt tctcccccac cctcgggaaa aaggcggagc cagtacacga catcactttc
     4861 ccagtttacc ccgcgccacc ttctctaggc accggttcaa ttgccgaccc ctccccccaa
     4921 cttctcgggg actgtgggcg atgtgcgctc tgcccactga cgggcaccgg agccaattcc
     4981 cactcctttc aagacctaga aggtccatta gctgcaaaga ttcctctctg tttaaaactt
     5041 tatccatctt tgcaaagctt tttgcaaaag cctagtcaat aatcaatgtc aacgcgtata
     5101 tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca
     5161 attgtcggtc aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac
     5221 ctccaacaca caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt
     5281 actgagagtg caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact
     5341 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5401 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5461 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5521 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5581 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5641 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5701 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5761 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5821 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5881 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5941 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6001 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6061 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6121 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6181 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6241 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6301 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6361 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6421 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6481 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6541 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6601 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6661 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6721 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6781 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6841 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6901 cttcccgaag ggagaaaggc ggac
//
