LOCUS       sequence-349066-        7536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single INBox (INCENP 819-918) recruitment responsible for targeted 
            Aurora B activation and recruitment using TMP-Halo dimerization 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7536)
  AUTHORS   Chen GY, Renda F, Zhang H, Gokden A, Wu DZ, Chenoweth DM, Khodjakov 
            A, Lampson MA
  TITLE     Tension promotes kinetochore-microtubule release by Aurora B kinase.
  JOURNAL   J Cell Biol. 2021 Jun 7;220(6). pii: 212027. doi: 
            10.1083/jcb.202007030.
  PUBMED    33904910
REFERENCE   2  (bases 1 to 7536)
  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: "J Cell 
            Biol."; date: "2021-06-7"; volume: "220(6). pii: 212027. doi"; 
            pages: " 10.1083/jcb.202007030"
COMMENT     Sequence Label: CAG.mCherry-INBox-eDHFR
FEATURES             Location/Qualifiers
     source          1..7536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          127..1143
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1067..1089
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1151..1170
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1226..1933
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1371..1389)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1639..1658
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2267..2740
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             2741..2770
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2877..2896)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2942..2997
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2996..3015)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    3361..3394
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3407..3528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3444..3463)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3498..3517
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3547..3567)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3547..3565)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3586..3602)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3586..3602)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3599..3621)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3610..3626
                     /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)."
     promoter        complement(join(3634..3640,3641..3658,3659..3664))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3679..3700
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3817..3834)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3988..4576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4068..4087)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4747..5538,5539..5607))
                     /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     5370..5389
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5608..5712)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5785..5833
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    6045..6078
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             6083..6682
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6083..6102)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6579..6599
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6818..6835)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6824..6935
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        7080..7383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7385..7536,1..126)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
       61 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      121 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      181 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      241 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      301 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      361 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      421 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
      481 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
      541 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
      601 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
      661 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
      721 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
      781 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
      841 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
      901 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
      961 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1021 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1081 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1141 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgctc
     1201 gaccaccggc gcgccaccgg tcaccatggt gagcaagggc gaggaggata acatggccat
     1261 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     1321 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     1381 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     1441 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     1501 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     1561 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     1621 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     1681 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     1741 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     1801 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     1861 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     1921 cgagctgtat aagggtaccc cagataacta cgggatggat ctgaatagcg acgactccac
     1981 cgatgatgag gcccatcccc ggaagcccat ccccacctgg gcccgaggca ccccgctcag
     2041 ccaggctatc attcaccagt actaccaccc accgaacctt ctggagctct ttggaaccat
     2101 tctcccactg gacttggagg atatcttcaa gaagagcaag ccccgctatc acaagcgcac
     2161 cagctctgct gtctggaact caccgcccct gcagggcgcc agggtcccca gcagcctggc
     2221 ctacagcctg aagaagcacg gatccaccgg atccctggag tctagaatca gtctgattgc
     2281 ggcgttagcg gtagatcgcg ttatcggcat ggaaaacgcc atgccgtgga acctgcctgc
     2341 cgatctcgcc tggtttaaac gcaacacctt aaataaaccc gtgattatgg gccgccatac
     2401 ctgggaatca atcggtcgtc cgttgccagg acgcaaaaat attatcctca gcagtcaacc
     2461 gggtacggac gatcgcgtaa cgtgggtgaa gtcggtggat gaagccatcg cggcgtgtgg
     2521 tgacgtacca gaaatcatgg tgattggcgg cggtcgcgtt tatgaacagt tcttgccaaa
     2581 agcgcaaaaa ctgtatctga cgcatatcga cgcagaagtg gaaggcgaca cccatttccc
     2641 ggattacgag ccggatgact gggaatcggt attcagcgaa ttccacgatg ctgatgcgca
     2701 gaactctcac agctattgct ttgagattct ggagcggcgg gaacaaaaac tcatctcaga
     2761 agaggatctt taattaagcg ctcgaggcgg ccgcaattca ctcctcaggt gcaggctgcc
     2821 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     2881 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     2941 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3001 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3061 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     3121 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     3181 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     3241 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     3301 gtccctcttc tcttatggag atccctcgac ctgcaccgtc gagctagcat atggatccat
     3361 ataacttcgt ataatgtatg ctatacgaag ttattacgat accgtcaact tgtttattgc
     3421 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3481 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttagg tacccagctt
     3541 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     3601 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3661 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3721 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3781 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3841 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3901 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3961 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4021 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4081 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4141 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4201 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4261 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4321 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4381 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4441 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4501 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4561 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4621 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4681 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4741 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4801 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4861 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4921 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4981 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5041 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5101 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5161 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5221 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5281 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5341 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5401 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5461 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5521 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5581 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5641 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5701 aggggttccg cgcacatttc cccgaaaagt gccaagctta ataaacacgg aaggagacaa
     5761 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     5821 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5881 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5941 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     6001 actggccccg tgggttaggg acggggtccc ccattcgaga ataaataact tcgtataaag
     6061 tatcctatac gaagttatca ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     6121 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     6181 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     6241 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     6301 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     6361 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     6421 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     6481 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     6541 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     6601 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     6661 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     6721 cgcacgaccc catggctccg accgaagcca cccggggcgg ccccgccgac cccgcacccg
     6781 cccccgaggc ccaccgactc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     6841 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     6901 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     6961 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcact
     7021 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7081 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7141 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7201 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7261 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7321 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7381 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     7441 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
     7501 ggggggcgcg cgccaggcgg ggcggggcgg ggcgag
//
