LOCUS       sequence-430214-        5030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human H4 carrying three point mutations to disrupt 
            binding with H3. Expression in mammalian cells driven by the CMV 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5030)
  AUTHORS   Pardal AJ, Bowman AJ
  TITLE     A specific role for importin-5 and NASP in the import and nuclear 
            hand-off of monomeric H3.
  JOURNAL   Elife. 2022 Sep 6;11:e81755. doi: 10.7554/eLife.81755.
  PUBMED    36066346
REFERENCE   2  (bases 1 to 5030)
  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: "Elife."; 
            date: "2022-09-6"; volume: "11:e81755. doi"; pages: " 
            10.7554/eLife.81755"
COMMENT     Sequence Label: pCL1318
FEATURES             Location/Qualifiers
     source          1..5030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      115..570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(202..221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     412..433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    577..698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(588..607)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     642..661
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(821..1534,1535..1537))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITLGMDELYK"
     primer_bind     1215..1234
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1474..1495
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1948..2151)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1977..1997)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2152..2455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2630..3218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2710..2729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3547..3594
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3619..3638
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3826..4620)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3826..4620)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3937..3956)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4547..4566
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4655..5012)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4669..4804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4821..5012
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4969..4989
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(join(5014..5030,1..88))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
       61 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
      121 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      181 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      241 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      301 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
      361 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      421 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      481 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      541 attttaacaa aatattaacg cttacaattt acgccttaag atacattgat gagtttggac
      601 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      661 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      721 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      781 aatgtggtat ggctgattat gatctagagt cgcggccgct ttacttgtac agctcgtcca
      841 tgccgagagt gatcccggcg gcggtcacga actccttaag gaccatgtga tcgcgcttct
      901 cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca
      961 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     1021 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     1081 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     1141 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     1201 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     1261 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     1321 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
     1381 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     1441 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     1501 gcaccacccc ggtgaacagc tcctcgccct tgctcaccgt gctggcgacc ggtagcgcac
     1561 ctccacctga gccacctcca gatccacctc ctgaaccacc tccgcctccg aagccgtaca
     1621 gggtacggcc ctgacgtttc agagcgtaaa caacgtccat agcggtaacg gttttacgtt
     1681 tagcgtgttc ggtgtaggta acagcgtcac gtgcaacgtt ttccagagca actttagcaa
     1741 caccgcgggt ttcttcgtag atcagaccgg agatacgttt aacaccacca cgacgagcca
     1801 gacgacggat agccggcttg gtgataccct ggatgttgtc acgcagaact ttacggtgac
     1861 gtttagcacc acctttaccc agacctttac cgccctttcc tctgccggac atggcggcgg
     1921 ctagcggatc tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg
     1981 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2041 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2101 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2161 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2221 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2281 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2341 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2401 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     2461 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca
     2521 tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2581 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2641 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2701 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2761 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2821 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2881 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2941 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3001 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3061 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3121 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3181 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3241 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3301 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3361 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     3421 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     3481 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     3541 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     3601 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     3661 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     3721 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     3781 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     3841 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     3901 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     3961 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     4021 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     4081 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     4141 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     4201 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     4261 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     4321 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     4381 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     4441 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     4501 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     4561 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     4621 gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa
     4681 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     4741 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     4801 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     4861 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     4921 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     4981 acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat
//
