LOCUS       sequence-172552-        5810 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  acute activation of Rho through rapamycin-induced translocation to 
            membrane-localized FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5810)
  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 5810)
  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(ARHGEF1)
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     256..278
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     340..359
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             393..1100
                     /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(538..556)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     806..825
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1113..1433
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(2353..2372)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2418..2473
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2472..2491)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2834..2850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2834..2850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2847..2869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2858..2874
                     /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(2882..2888,2889..2906,2907..2912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2927..2948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3007..3202
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3053..3188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3208..3342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3245..3264)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3299..3318
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3410..3427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3581..4169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3661..3680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4340..5131,5132..5200))
                     /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     4963..4982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5201..5305)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5336..5715
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
       61 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
      121 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
      181 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
      241 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
      301 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
      361 tgtctcatca ttttggcaaa gaattcaccg gtatggtgag caagggcgag gaggataaca
      421 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
      481 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      541 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
      601 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
      661 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
      721 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
      781 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
      841 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
      901 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
      961 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     1021 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     1081 gcatggacga gctgtacaag agtgctggtg gtggagtgca ggtggaaacc atctccccag
     1141 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     1201 ttgaagatgg aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc
     1261 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     1321 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     1381 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaaagtgctg
     1441 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtcctcgag
     1501 gtgcggccgc cgagggggag ccggggcggt cgggactgga gcttgaacca gaagagcctc
     1561 ccggctggcg ggaactcgtc cccccagaca ccctgcacag cctgcccaag agccaggtga
     1621 agcggcagga ggtcatcagc gagctgctgg tgacagaggc ggcccacgtg cgcatgctgc
     1681 gggtgctgca cgacctcttc ttccagccca tggcagaatg cctgttcttc cccttggagg
     1741 agctgcagaa catcttcccc agcctggacg agctcatcga ggtgcattcc ctgttcctcg
     1801 atcgcctgat gaagcggagg caggagagtg gctacctcat cggggagatc ggagacgtgc
     1861 tgctggcccg gtttgatggt gctgagggct cctggttcca gaaaatctcc tcccgcttct
     1921 gcagccgcca gtcatttgcc ttagagcagc tcaaagccaa gcaacgcaag gaccctcggt
     1981 tctgtgcctt cgtgcaggaa gctgagagcc gcccgcggtg ccgccgcctg cagctgaagg
     2041 acatgatccc cacggagatg cagcggctga ccaagtaccc cctgctcctg cagagcatcg
     2101 ggcagaacac agaagcgccc acagaacggg agaaagtgga gctggcagcc gagtgctgcc
     2161 gggaaattct acaccacgtc aaccaagccg tgcgtgacat ggaggacctg ctgaggctca
     2221 aggactatca gcggcgcctg gacttgtccc accttcggca gagctgagcg gccgcactcc
     2281 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     2341 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     2401 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2461 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     2521 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     2581 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     2641 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     2701 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     2761 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     2821 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2881 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2941 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3001 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3121 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3181 tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa
     3241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3301 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg
     3361 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4021 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4261 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4321 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4381 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4441 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4501 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4561 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4621 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4681 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4741 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4801 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4861 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4921 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4981 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5041 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5101 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5161 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5281 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
     5341 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     5401 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5461 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5521 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5581 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     5641 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     5701 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     5761 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
//
