LOCUS       sequence-419565-        5111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning of GFP-H2B fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5111)
  AUTHORS   Pfisterer M, Robert R, Saul VV, Pritz A, Seibert M, Feederle R, 
            Schmitz ML
  TITLE     The Aurora B-controlled PP1/RepoMan complex determines the spatial 
            and temporal distribution of mitotic H2B S6 phosphorylation.
  JOURNAL   Open Biol. 2024 May;14(5):230460. doi: 10.1098/rsob.230460. Epub 
            2024 May 29.
  PUBMED    38806145
REFERENCE   2  (bases 1 to 5111)
  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: "Open 
            Biol."; date: "2024-05"; volume: "14(5):230460. doi"; pages: " 
            10.1098/rsob.230460. Epub 2024 May 29"
COMMENT     Sequence Label: pEGFP-H2B-MCS
FEATURES             Location/Qualifiers
     source          1..5111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             597..974
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      987..996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(993..995,996..998,999..1709)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(993..995,996..998,999..1709)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1038..1059)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1299..1318)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1646..1667
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1710..1775
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1899..2020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1936..1955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1990..2009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2027..2482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2164..2185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2376..2395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2509..2613
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2615..2972
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2615..2806
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2638..2658)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2823..2958
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3007..3801
                     /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             3007..3801
                     /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(3061..3080)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3671..3690
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3989..4008)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4033..4080
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4409..4997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4898..4917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgc
      601 cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg
      661 cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg
      721 tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca
      781 tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg
      841 cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc
      901 tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt
      961 acaccagcgc taaggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     1021 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1081 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1141 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1201 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1261 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1321 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1381 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1441 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1501 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1561 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1621 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1681 tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga
     1741 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     1801 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1861 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1921 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1981 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2041 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2101 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2161 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2221 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2281 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2341 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2401 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2461 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2521 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2581 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2641 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2701 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2761 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2821 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2881 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2941 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3001 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3061 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3121 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3181 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3241 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3301 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3361 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3421 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3481 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3541 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3601 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3661 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3721 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3781 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3841 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3901 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3961 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4021 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4081 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4141 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4201 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4261 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4321 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4381 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4441 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4501 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4561 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4621 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4681 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4741 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4801 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4861 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4921 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4981 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5041 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5101 ccgccatgca t
//
