LOCUS       sequence-386848-        5190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human CENP-B (aa 1-169 containing DNA-binding domain) cloned into 
            pDsRed1-N1 vector (Clontech)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5190)
  AUTHORS   Chagin VO, Reinhart B, Becker A, Mortusewicz O, Jost KL, Rapp A, 
            Leonhardt H, Cardoso MC
  TITLE     Processive DNA synthesis is associated with localized decompaction 
            of constitutive heterochromatin at the sites of DNA replication and 
            repair.
  JOURNAL   Nucleus. 2019 Dec;10(1):231-253. doi: 10.1080/19491034.2019.1688932.
  PUBMED    31744372
REFERENCE   2  (bases 1 to 5190)
  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"; doi: 
            "10.1080/19491034.2019"; journalName: "Nucleus"; date: "2019-12"; 
            volume: "10"; issue: "1"; pages: "231-253"
COMMENT     Sequence Label: pCENPBR pc861
FEATURES             Location/Qualifiers
     source          1..5190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1183..1192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1189..1191,1192..1194,1195..1869)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1372..1392)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1590..1609
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1784..1807
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1990..2111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2027..2046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2081..2100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2118..2573)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2255..2276)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2467..2486
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2600..2704
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2706..3063
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2706..2897
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2729..2749)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2914..3049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3098..3892
                     /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             3098..3892
                     /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(3152..3171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3762..3781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4080..4099)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4124..4171
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4500..5088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4989..5008
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagcgc taccggactc agatctcgag ctcaagcttg gtaccgggcg cgccgggatg
      661 ggccccaagc ggcggcagct gacgttccgg gagaagtcgc ggatcatcca ggaggtggag
      721 gagaacccgg acctacgcaa gggcgagatc gcgcggcgct tcaacatccc gccgtccacg
      781 ctgagcacga tcctgaagaa caagcgcgcc atcctggcgt cggagcgcaa gtacggggtg
      841 gcctccacct gccgcaagac caacaagctg tctccctacg acaagctcga gggcttgctc
      901 atcgcctggt tccagcagat ccgcgccgcc ggcctgccgg tcaagggcat catcctcaag
      961 gagaaggcgc tgcgcatagc cgaggagctg ggcatggacg acttcaccgc ctccaacggc
     1021 tggctggacc gcttccgccg gcgccacggc gtggtgtcct gcagcggcgt ggcccgcgcc
     1081 cgcgcgcgaa acgctgcccc ccgcaccccg gcggcgcctg ccagtccggc cgcggtgccc
     1141 tcggagggca gtggcgggag cactggtcta gatccaccgg tcgccaccat ggtgcgctcc
     1201 tccaagaacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg caccgtgaac
     1261 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg ccacaacacc
     1321 gtgaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     1381 cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag
     1441 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     1501 gtggtgaccg tgacccagga ctcctccctg caggacggct gcttcatcta caaggtgaag
     1561 ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     1621 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccacaag
     1681 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagtccat ctacatggcc
     1741 aagaagcccg tgcagctgcc cggctactac tacgtggact ccaagctgga catcacctcc
     1801 cacaacgagg actacaccat cgtggagcag tacgagcgca ccgagggccg ccaccacctg
     1861 ttcctgtagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     1921 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1981 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2041 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2101 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2161 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2221 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2281 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2341 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2401 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2461 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2521 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2581 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2641 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2701 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2821 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2881 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2941 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3001 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3061 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3121 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3181 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3241 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3301 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3361 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3421 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3481 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3541 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3601 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     3661 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3721 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3781 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3841 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3901 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3961 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4021 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4081 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4141 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4201 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4261 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4321 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4381 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4441 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4501 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4561 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4621 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4681 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4741 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4801 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4861 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4921 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4981 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5041 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5101 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5161 ttatcccctg attctgtgga taaccgtatt
//
