LOCUS       sequence-430209-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human H3.1 tagged on its C-terminus with eGFP. 
            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: pCL1313
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     307..326
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     566..587
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1100..1109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1141..1344)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1170..1190)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1345..1648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1823..2411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1903..1922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2740..2787
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2812..2831
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3019..3813)
                     /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(3019..3813)
                     /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(3130..3149)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3740..3759
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3848..4205)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3862..3997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4014..4205
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4162..4182
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4207..4311)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4338..4793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4425..4444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4635..4656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4800..4921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4811..4830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4865..4884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5044..5131,1..626,627..629))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITLGMDELYK"
ORIGIN
        1 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
       61 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
      121 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
      181 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
      241 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
      301 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
      361 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
      421 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
      481 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
      541 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
      601 ccggtgaaca gctcctcgcc cttgctcacc gtgctggcga ccggtagcgc gcctccacct
      661 gagccacctc ctgagccacc tcctgagcca cctccagctc tctctccgcg gatgcggcgg
      721 gcgagctgga tgtccttggg catgatagtg acgcgcttgg cgtggatggc gcacaggtta
      781 gtgtcctcaa atagccctac caagtaggcc tcgcacgcct cctgcagagc catcacagcg
      841 gagctctgga aacgcaggtc tgttttaaag tcctgcgcaa tctcgcgcac caggcgctgg
      901 aaaggtagtt tacgaataag cagttcagtg gacttctgat aacggcggat ctcgcgcaga
      961 gccacggtgc ccggccggta gcggtggggc tttttcacgc cgccggtggc cggagcgctt
     1021 ttgcgggctg ccttagtggc caactgtttg cgtggcgcct tgccaccagt agacttccga
     1081 gcagtttgct tagtgcgagc catggcggcg gcgctagcgg atctgacggt tcactaaacc
     1141 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     1201 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     1261 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     1321 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     1381 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     1441 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     1501 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     1561 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     1621 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     1681 ccccgtaatt gattactatt aataactaat gcatggcggt aatacggtta tccacagaat
     1741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2341 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg
     2581 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     2641 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     2701 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt
     2761 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
     2821 tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc
     2881 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     2941 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     3001 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3061 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     3121 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     3181 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     3241 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
     3301 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     3361 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     3421 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     3481 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     3541 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     3601 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     3661 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     3721 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     3781 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     3841 atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     3901 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     3961 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     4021 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     4081 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     4141 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     4201 ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4261 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4321 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4741 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttacgcctt
     4801 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4861 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4921 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4981 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc
     5041 gctttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactcctt
     5101 aaggaccatg tgatcgcgct tctcgttggg g
//
