LOCUS       sequence-261243-        9075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express a TALE construct that recognizes major satellite repeats 
            fused to mClover and the kinase domain of mouse BUB1 (aa 672-1058) 
            at the C terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9075)
  AUTHORS   Akera T, Trimm E, Lampson MA
  TITLE     Molecular Strategies of Meiotic Cheating by Selfish Centromeres.
  JOURNAL   Cell. 2019 Aug 22;178(5):1132-1144.e10. doi: 
            10.1016/j.cell.2019.07.001. Epub 2019 Aug 8.
  PUBMED    31402175
REFERENCE   2  (bases 1 to 9075)
  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: 
            "10.1016/j.cell.2019.07.001"; journalName: "Cell"; date: 
            "2019-08-22- 22"; volume: "178"; issue: "5"; pages: "1132-1144.e10"
COMMENT     Sequence Label: Major Sat-BUB1
FEATURES             Location/Qualifiers
     source          1..9075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    50..71
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(86..91,92..109,110..116)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    124..140
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     129..151
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     148..164
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     148..164
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        184..563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        565..842
                     /label=chicken beta-actin promoter
     intron          843..1859
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1783..1805
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1872..1891
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1872..1890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1910..1930
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4061..4063,4064..4066,4067..4777)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(4106..4127)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4714..4735
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6149..6178
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6194..6211
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6234..6251)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6240..6464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6474..6491)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6474..6490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6483..6505)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6632..7060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6719..6738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6929..6950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7113..7217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7218..7286,7287..8078)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7436..7455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8249..8837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8738..8757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8991..9008
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
       61 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      121 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgag gcgcgccgga
      181 ttcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      241 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      301 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      361 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      421 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      481 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      541 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      601 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      661 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggcggggcgg ggcgrggggc
      721 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      781 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      841 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      901 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      961 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
     1021 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
     1081 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
     1141 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1201 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1261 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1321 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1381 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1441 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1501 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1561 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1621 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1681 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1741 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1801 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacaga
     1861 tccttaatta ataatacgac tcactatagg ggccgccacc atgagatctc ctaagaaaaa
     1921 gaggaaggtg caggtggatc tacgcacgct cggctacagt cagcagcagc aagagaagat
     1981 caaaccgaag gtgcgttcga cagtggcgca gcaccacgag gcactggtgg gccatgggtt
     2041 tacacacgcg cacatcgttg cgctcagcca acacccggca gcgttaggga ccgtcgctgt
     2101 cacgtatcag cacataatca cggcgttgcc agaggcgaca cacgaagaca tcgttggcgt
     2161 cggcaaacag tggtccggcg cacgcgccct ggaggccttg ctcacggatg cgggggagtt
     2221 gagaggtccg ccgttacagt tggacacagg ccaacttgtg aagattgcaa aacgtggcgg
     2281 cgtgaccgca atggaggcag tgcatgcatc gcgcaatgca ctgacgggtg cccccctgaa
     2341 cctgaccccg gaccaagtgg tggctatcgc cagcaacggt ggcggcaagc aagcgctcga
     2401 aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt
     2461 ggtggctatc gccagcaaca agggcggcaa gcaagcgctc gaaacggtgc agcggctgtt
     2521 gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca
     2581 cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga
     2641 ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc
     2701 gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga
     2761 ccaagtggtg gctatcgcca gcaacattgg cggcaagcaa gcgctcgaaa cggtgcagcg
     2821 gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc
     2881 cagcaacggt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg
     2941 ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa
     3001 gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac
     3061 cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt
     3121 gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc
     3181 tatcgccagc aacggtggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt
     3241 gctgtgccag gaccatggcc tgactccgga ccaagtggtg gctatcgcca gccacgatgg
     3301 cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg
     3361 cctgaccccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga
     3421 aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt
     3481 ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt
     3541 gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca
     3601 cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga
     3661 ccatggcctg accccggacc aagtggtggc tatcgccagc aacaagggcg gcaagcaagc
     3721 gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga
     3781 ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa gcattgtggc
     3841 ccagctgagc cggcctgatc cggcgttggc cgcgttgacc aacgaccacc tcgtcgcctt
     3901 ggcctgcctc ggcggacgtc ctgccatgga tgcagtgaaa aagggattgc cgcacgcgcc
     3961 ggaattgatc agaagagtca atcgccgtat tggcgaacgc acgtcccatc gcgttgcggg
     4021 atccactgga ggcggaggtg gctccggcac cgcctcctcc atggtgagca agggcgagga
     4081 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     4141 gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     4201 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
     4261 cggcgtggcc tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4321 cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg acggtaccta
     4381 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4441 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     4501 cagccacaac gtctatatca cggccgacaa gcagaagaac ggcatcaagg ctaacttcaa
     4561 gatccgccac aacgttgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4621 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc atcagtccaa
     4681 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4741 cgccgggatt acacatggca tggacgagct gtacaagggt ggtgaggctt ttaaatgcac
     4801 aggcattgac catgcgacag tggaagacct atccgatgcc aatgctgggc tccaagttga
     4861 atgcgtgcag acacttggaa atgtcaatgc tccaagcttt actgttgaga acccatggga
     4921 tgatgaattg attcttaaac ttctctctgg actttctaag ccagttactt cctattcaaa
     4981 tacttttgag tggcagagta aacttccagc catcaagacc aagacagaat atcaattggg
     5041 ttctttgctg gtctatgtga atcaccttct tggagaagga gcctttgctc aagtctttga
     5101 agctattcat ggagatgtga gaaatgccaa aagtgaacag aaatgcattt tgaaggtgca
     5161 gagacctgcc aactcctggg aattctacat tgggatgcag ctgatggaaa gactaaagcc
     5221 agaagtacat cacatgttca tcaagtttta ttctgctcat ttattcaaga acggcagcat
     5281 attagtaggg gaactctaca gctatgggac gttactaaat gtcattaacc tctataaaaa
     5341 tacctctgaa aaagtgatgc cccaggctct tgtcctcact ttcgctatca gaatgcttta
     5401 catggttgaa caagtccaca gctgcgaaat cattcatgga gacattaagc cagataactt
     5461 catactagga cacagatttt tggaacaggc tgatgaagac ttagctaccg gcttggcatt
     5521 gattgacctg ggtcagagta tagatatgaa acttttccct aaaggaactg tatttacagg
     5581 aaaatgtgaa acatctggtt ttcagtgtcc tgagatgctc agtaacaagc catggaacta
     5641 ccagattgat tactttggag ttgctgcaac aatatactgt atgctctttg gctcttacat
     5701 gaaagtaaaa aatgaaggag gagtctggaa acctgaaggt ctttttagaa ggcttcctca
     5761 tttggatatg tgggaggaat tttttcacat catgttgaat ataccggatt gtcataatct
     5821 tccatctttg gattttctga gacagaatat gaagaaatta cttgaacaac agtattccaa
     5881 caagattaag accttgcgta ataggctaat tgtgatgctt tcagaatata agcgttcaag
     5941 aaaataacct aggcttaagg tacatatggg caccaaagaa cctgtaaacg ttatcttttt
     6001 aaattgaatg tgcacaaata aaagwwwrga aaagaaaaaa aaaaaaaaaa aaaaaaaaaa
     6061 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaatacgcat catacgctgt
     6121 ggcgtacctc gagtctagag ggcccttcga acaaaaactc atctcagaag aggatctgaa
     6181 tatgcatacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     6241 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6301 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6361 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6421 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atgggcggcc gccactggcc
     6481 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     6541 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     6601 caacagttgc gcagcctgaa tgksgmaysk ramkygycct gtagcggcgc attaagcgcg
     6661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6781 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6841 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6901 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6961 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7021 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacsmkk
     7081 rcrmktymsr tkssmcwtty mggsgaaatg tgcgcggaac ccctatttgt ttatttttct
     7141 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7201 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7261 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7321 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7381 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7441 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7501 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7561 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     7621 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     7681 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7741 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7801 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7861 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7921 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7981 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8041 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8101 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8161 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8221 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     8281 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8341 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     8401 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     8461 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     8521 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     8581 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     8641 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     8701 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     8761 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     8821 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     8881 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     8941 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     9001 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     9061 ttcattaatg cagct
//
