LOCUS       sequence-430212-        5030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human H4 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 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: pCL1316
FEATURES             Location/Qualifiers
     source          1..5030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..382
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        25..216
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(48..68)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      233..368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             417..1211
                     /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             417..1211
                     /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(471..490)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1081..1100
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1399..1418)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1443..1490
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1819..2407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2308..2327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2582..2885
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2886..3089
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3040..3060
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(3500..3502,3503..4216)
                     /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(3542..3563)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3803..3822)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4339..4460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4376..4395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4430..4449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4467..4922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4604..4625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4816..4835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4949..5030,1..23)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
       61 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      121 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      181 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      241 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      301 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
      361 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
      421 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
      481 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
      541 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
      601 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
      661 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
      721 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
      781 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
      841 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
      901 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
      961 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     1021 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     1081 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     1141 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     1201 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     1261 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1321 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     1381 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     1441 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     1501 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     1561 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     1621 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     1681 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1741 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1801 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1861 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1921 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1981 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2041 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2101 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2161 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2221 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2281 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2341 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2401 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2461 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     2521 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     2581 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     2641 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     2701 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     2761 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     2821 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     2881 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     2941 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     3001 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     3061 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagccgc
     3121 cgccatgtcc ggcagaggaa agggcggtaa aggtctgggt aaaggtggtg ctaaacgtca
     3181 ccgtaaagtt ctgcgtgaca acatccaggg tatcaccaag ccggctatcc gtcgtctggc
     3241 tcgtcgtggt ggtgttaaac gtatctccgg tctgatctac gaagaaaccc gcggtgttct
     3301 gaaagttttc ctggaaaacg ttatccgtga cgctgttacc tacaccgaac acgctaaacg
     3361 taaaaccgtt accgctatgg acgttgttta cgctctgaaa cgtcagggcc gtaccctgta
     3421 cggcttcgga ggcggaggtg gttcaggagg tggatctgga ggtggctcag gtggaggtgc
     3481 gctaccggtc gccagcacgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3541 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3601 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3661 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3721 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3781 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3841 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3901 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3961 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     4021 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4081 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     4141 gcgcgatcac atggtcctta aggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4201 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     4261 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4321 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4381 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4441 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     4501 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4561 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4621 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4681 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4741 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4801 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4861 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4921 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4981 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
//
