LOCUS       sequence-419560-        7717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses H2B-GFP fused to INCENP∆cen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7717)
  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 7717)
  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: H2B-GFP-INCENP∆cen
FEATURES             Location/Qualifiers
     source          1..7717
                     /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"
     polyA_signal    4505..4626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4542..4561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4596..4615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4633..5088)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4770..4791)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4982..5001
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5115..5219
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5221..5578
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5221..5412
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5244..5264)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5429..5564
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5613..6407
                     /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             5613..6407
                     /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(5667..5686)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6277..6296
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6595..6614)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6639..6686
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7015..7603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7504..7523
                     /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 atctcgagtc agcaaagagc
     1741 cagagctgat gcccaaaaca ccttctcaga agaaccgacg gaagaagaga cggatttctt
     1801 atgttcagga tgaaaacaga gatcccatca ggagaaggtt atcccgcaga aagtctcgga
     1861 gcagccagct gagctcccga cgcctccgca gcaaggacag tgtagagaag ctggctacag
     1921 tggtcgggga gaacggctcc gttctgcggc gtgtgacccg tgctgcggct gcagctgccg
     1981 cggctaccat ggcattggct gcaccttctt cacccacccc tgagtctccc acgatgctga
     2041 ctaagaagcc cgaggataac cacacccagt gccagctggt gcctgtggtg gagatcggca
     2101 tcagtgagcg ccagaatgct gagcagcatg tcacccagct catgtccacc gagcctctgc
     2161 cccgcactct gtccccgact ccagcttcag ccacagctcc aacctcccag ggcatcccga
     2221 catcagatga ggaatcaaca cctaagaagt cgaaggccag gatactggag tccatcacag
     2281 tgagctccct gatggctaca ccccaggacc ccaagggtca aggggtcggg acggggcggt
     2341 ctgcgtctaa gctcaggatt gcgcaggtct cccctggccc acgggactcg ccagcctttc
     2401 cagattctcc atggcgggag cgggtgctgg ctcccatcct gccggataac ttctccacgc
     2461 ccacgggctc tcgcacggac tctcaatcgg tgcggcacag cccgatcgcc ccgtcttccc
     2521 cgagtccccg agtcttagcc cagaagtact ctctggtggc caaacaggaa agtgttgtcc
     2581 gcagggcgag cagaaggctt gccaagaaga ctgccgaaga gccagctgcc tctggccgca
     2641 tcatctgtca cagttacctg gagaggctcc tgaatgttga ggtgccccag aaagttggtt
     2701 ctgagcagaa ggaacccccc gaggaggctg agcctgtggc ggcagctgag ccagaggtcc
     2761 ctgagaacaa cggaaataac tcgtggcccc acaatgacac agagatcgcc aacagcacac
     2821 ccaacccgaa gcctgcagcc agcagcccgg aaacaccctc tgcagggcag caagaggcca
     2881 agacggacca agcagatgga cccagagagc caccgcagag tgccaggagg aagcgcagct
     2941 acaagcaggc cgtgagtgag ctggacgagg agcagcacct ggaggatgag gagctgcagc
     3001 cccccaggag caagacccct tcctcaccct gcccagccag caaggtggta cggcccctcc
     3061 ggacctttct gcacacagtg cagaggaacc agatgctcac gaccccgacc tcagccccac
     3121 gcagcgtcat gaagtccttt attaagcgca acactcccct gcgcatggac cccaagtgca
     3181 gcttcgtcga gaaggagcgg cagcgcctgg agaatctgcg gcggaaggag gaggccgagc
     3241 agctgcgcag gcagaaggtg gaggaggaca agcggcggcg gctggaggag gtgaagctga
     3301 agcgtgagga acgcctccgc aaggtgctgc aggcccgcga gcgggtggag cagatgaagg
     3361 aggagaagaa gaagcagatt gagcagaagt ttgctcagat cgacgagaag actgagaagg
     3421 ccaaggagga gcggctggca gaggagaagg ccaagaaaaa ggcggcggcc aagaagatgg
     3481 aggaggtgga agcacgcagg aagcaggaag aggatgcacg taggctcagg tggctgcagc
     3541 aggaggagga agagcggcgg caccaagagc tgctgcagaa gaagaaggaa gaggagcagg
     3601 agcggctgcg gaaggcggcc gaggctaagc ggctggcaga gcagcgggag caggagcggc
     3661 gggagcagga gcggcgcgag caggagcggc gcgagcagga gcggcgggag caggagcggc
     3721 gcgagcagga gcgacagctg gcagagcagg agcgtcggcg ggagcaggag cggctccagg
     3781 ccgagaggga gctgcaggag cgggagaagg ccctgcggct gcagaaggag cagctgcaga
     3841 gggaactgga ggagaagaag aagaaggaag agcagcagcg tctggctgag cggcagctgc
     3901 aggaggagca agagaagaaa gccaaggagg cagcaggggc cagcaaggcc ctgaatgtga
     3961 ctgtggacgt gcagtctcca gcttgtacct catatcagat gactccgcaa gggcacaggg
     4021 cccctcccaa gatcaaccca gataactacg ggatggatct gaatagcgac gactccaccg
     4081 atgatgaggc ccatccccgg aagcccatcc ccacctgggc ccgaggcacc ccgctcagcc
     4141 aggctatcat tcaccagtac taccacccac cgaaccttct ggagctcttt ggaaccattc
     4201 tcccactgga cttggaggat atcttcaaga agagcaagcc ccgctatcac aagcgcacca
     4261 gctctgctgt ctggaactca ccgcccctgc agggcgccag ggtccccagc agcctggcct
     4321 acagcctgaa gaagtaaaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc
     4381 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4441 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4501 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4621 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4681 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4741 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4801 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4861 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4921 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4981 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5041 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     5101 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5161 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5221 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5281 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5341 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5401 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5461 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5521 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5581 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5641 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5701 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5761 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5821 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5881 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5941 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6001 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6061 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6121 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6181 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6241 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6301 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6361 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6421 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6481 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6541 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6601 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6661 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6721 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6781 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6841 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6901 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6961 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7021 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7081 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7141 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7201 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7261 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7321 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7381 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7441 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7501 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7561 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7621 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7681 ccctgattct gtggataacc gtattaccgc catgcat
//
