LOCUS       sequence-172553-        5850 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  acute activation of Rac through rapamycin-induced translocation to 
            membrane-localized FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5850)
  AUTHORS   Hayer A, Shao L, Chung M, Joubert LM, Yang HW, Tsai FC, Bisaria A, 
            Betzig E, Meyer T
  TITLE     Engulfed cadherin fingers are polarized junctional structures 
            between collectively migrating endothelial cells.
  JOURNAL   Nat Cell Biol. 2016 Dec;18(12):1311-1323. doi: 10.1038/ncb3438. Epub
            2016 Nov 14.
  PUBMED    27842057
REFERENCE   2  (bases 1 to 5850)
  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: "10.1038/ncb3438"; 
            journalName: "Nat Cell Biol"; date: "2016-12"; volume: "18"; issue: 
            "12"; pages: "1311-1323"
COMMENT     Sequence Label: pCAG-mCherry-FKBP-GEF(TIAM1)
FEATURES             Location/Qualifiers
     source          1..5850
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     319..341
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     403..422
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             456..1163
                     /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(601..619)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     869..888
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1176..1496
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(2388..2407)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2453..2508
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2507..2526)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2869..2885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2869..2885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2882..2904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2893..2909
                     /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(2917..2923,2924..2941,2942..2947))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2962..2983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3042..3237
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3088..3223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3243..3377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3280..3299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3334..3353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3445..3462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3616..4204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3696..3715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4375..5166,5167..5235))
                     /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     4998..5017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5236..5340)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5371..5750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
       61 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
      121 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
      181 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
      241 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
      301 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
      361 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
      421 tgctgtctca tcattttggc aaagaattca ccggtatggt gagcaagggc gaggaggata
      481 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
      541 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
      601 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
      661 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
      721 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
      781 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
      841 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
      901 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
      961 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     1021 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     1081 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     1141 gcggcatgga cgagctgtac aagagtgctg gtggtggagt gcaggtggaa accatctccc
     1201 caggagacgg gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga
     1261 tgcttgaaga tggaaagaaa tttgattcct cccgggacag aaacaagccc tttaagttta
     1321 tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg
     1381 gtcagagagc caaactgact atatctccag attatgccta tggtgccact gggcacccag
     1441 gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaaagtg
     1501 ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggtcctc
     1561 gaggtgcggc cgccatgaac ccctctgacc agagcccatc tcctcaggac tccacggggc
     1621 ctcagctggc gaccatgaga caactctcgg atgcagataa gctgcgcaag gtgatctgcg
     1681 agctcctgga gacggagcgc acctacgtga aggatttaaa ctgtcttatg gagagatacc
     1741 taaagcctct tcaaaaagaa acttttctca cccaggatga gcttgacgtg ctttttggaa
     1801 atttaacgga aatggtagag tttcaagtag aattccttaa aactctagaa gatggagtga
     1861 gactggtacc tgatttggaa aagcttgaga aggttgatca atttaagaaa gtgctgttct
     1921 ctctgggggg atcattcctg tattatgctg accgcttcaa gctctacagt gccttctgcg
     1981 ccagccacac aaaagttccc aaggtcctgg tgaaagccaa gacagacacg gctttcaagg
     2041 cattcttgga tgcccagaac ccgaagcagc agcactcatc cacgctggag tcgtacctca
     2101 tcaagcccat ccagaggatc ctcaagtacc cacttctgct cagggagctg ttcgccctga
     2161 ccgatgcgga gagcgaggag cactaccacc tggacgtggc catcaagacc atgaacaagg
     2221 ttgccagtca catcaatgag atgcagaaaa tccatgaaga gtttggggct gtgtttgacc
     2281 agctgattgc ttgacctcga gggcggccgc actcctcagg tgcaggctgc ctatcagaag
     2341 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     2401 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     2461 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     2521 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     2581 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     2641 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     2701 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     2761 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     2821 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     2881 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2941 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3001 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     3061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3241 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3361 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3421 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3481 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3541 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3781 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4021 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4141 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4201 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4261 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4321 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4381 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4441 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4501 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4561 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4621 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4681 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4741 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4801 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4861 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4921 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4981 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5041 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5101 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5161 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5221 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5281 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5341 cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactagt tattaatagt
     5401 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     5461 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     5521 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     5581 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     5641 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     5701 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
     5761 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
     5821 tatttatttt ttaattattt tgtgcagcga
//
