LOCUS       sequence-430211-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human H3.1 carrying a triple point mutation to disrupt
            folding with histone H4. 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 5131)
  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 5131)
  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: pCL1315
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     403..422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(721..740)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    765..812
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1141..1729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1630..1649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1904..2207
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2208..2411
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2362..2382
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2443..2452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2923..2925,2926..3639)
                     /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     complement(2965..2986)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3226..3245)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3762..3883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3799..3818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3853..3872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3890..4345)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4027..4048)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4239..4258
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4372..4476
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4478..4835
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4478..4669
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4501..4521)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4686..4821
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4870..5131,1..533)
                     /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             join(4870..5131,1..533)
                     /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(4924..4943)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
       61 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      121 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      181 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
      241 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
      301 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
      361 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
      421 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
      481 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
      541 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
      601 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
      661 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
      721 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
      781 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
      841 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
      901 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
      961 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     1021 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1081 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1261 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1801 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     1861 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     1921 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1981 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2041 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2101 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2161 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     2221 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     2281 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2341 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2401 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ccgccgccat ggctcgcact
     2461 aagcaaactg ctcggaagtc tactggtggc aaggcgccac gcaaacagtt ggccactaag
     2521 gcagcccgca aaagcgctcc ggccaccggc ggcgtgaaaa agccccaccg ctaccggccg
     2581 ggcaccgtgg ctctgcgcga gatccgccgt tatcagaagt ccactgaact gcttattcgt
     2641 aaactacctt tccagcgcct ggtgcgcgag attgcgcagg actttaaaac agacctgcgt
     2701 gcacagagct ccgctgtgat ggctgcacag gaggcgtgcg aggccgcatt ggtagggcta
     2761 tttgaggaca ctaacctgtg cgccatccac gccaagcgcg tcactatcat gcccaaggac
     2821 atccagctcg cccgccgcat ccgcggagag agagctggag gtggctcagg aggtggctca
     2881 ggaggtggct caggtggagg cgcgctaccg gtcgccagca cggtgagcaa gggcgaggag
     2941 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3001 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3061 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     3121 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3181 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3241 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3301 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3361 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     3421 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3481 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     3541 ctgagcaaag accccaacga gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc
     3601 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     3661 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3721 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3781 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3841 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3901 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3961 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4021 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4081 ctaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4141 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4201 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4261 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4321 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4381 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4441 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4501 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4561 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4621 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4681 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4741 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4801 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4861 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4921 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4981 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5041 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5101 gcagctgtgc tcgacgttgt cactgaagcg g
//
