LOCUS       sequence-430213-        5044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human H4 carrying a triple glycine insertion to 
            disrupt folding 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 5044)
  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 5044)
  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: pCL1317
FEATURES             Location/Qualifiers
     source          1..5044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(62..81)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     272..293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    437..558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(448..467)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     502..521
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(681..1394,1395..1397))
                     /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     1075..1094
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1334..1355
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1822..2025)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1851..1871)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2026..2329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2504..3092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2584..2603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3421..3468
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3493..3512
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3700..4494)
                     /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(3700..4494)
                     /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(3811..3830)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4421..4440
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4529..4886)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4543..4678
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4695..4886
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4843..4863
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4888..4992)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5019..5044,1..430)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
       61 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
      121 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
      181 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
      241 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
      301 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
      361 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
      421 cttacaattt acgccttaag atacattgat gagtttggac aaaccacaac tagaatgcag
      481 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
      541 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
      601 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
      661 gatctagagt cgcggccgct ttacttgtac agctcgtcca tgccgagagt gatcccggcg
      721 gcggtcacga actccttaag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
      781 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
      841 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
      901 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
      961 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1021 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1081 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1141 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg
     1201 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     1261 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     1321 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1381 tcctcgccct tgctcaccgt gctggcgacc ggtagcgctc ctcctcctga gccacctcct
     1441 gaaccacctc ctgacccacc tccgcctccg aagccgtaca gggtgcgccc ctggcgcttg
     1501 agcgcgtaca ccacatccat ggctgtgacg gtcttgcgct tggcgtgctc ggtgtaggtg
     1561 actgcgtccc gaattcctcc acccacattc tccaggaaca ccttcagcac accgcgggtc
     1621 tcctcgtaaa tgaggccaga gatccgctta acgccgccac gccgagctag acgccgaatg
     1681 gcaggcttgg tgatgccctg aatgttgtct ctcaagacct tgcggtggcg cttagcgccc
     1741 cctttgccta agccttttcc gccctttcct ctgccggaca tcatggcggc ggcgctagcg
     1801 gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc
     1861 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     1921 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     1981 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat
     2041 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     2101 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     2161 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     2221 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     2281 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     2341 atatgggcta tgaactaatg accccgtaat tgattactat taataactaa tgcatggcgg
     2401 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2461 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2521 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2581 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2641 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2701 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2761 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2821 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2881 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2941 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3001 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3061 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3121 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3181 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3241 atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc
     3301 cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc
     3361 aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat
     3421 gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc atagcgcggg
     3481 ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg tgggcgaaga
     3541 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     3601 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
     3661 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag
     3721 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     3781 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     3841 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     3901 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     3961 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     4021 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     4081 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     4141 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     4201 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     4261 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc
     4321 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     4381 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     4441 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     4501 cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc
     4561 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     4621 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     4681 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     4741 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     4801 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     4861 tccacagctg gttctttccg cctcaggact cttccttttt caatattatt gaagcattta
     4921 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4981 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     5041 aagc
//
