LOCUS       sequence-166559-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  acute activation of Rac through rapamycin-induced translocation to 
            membrane-localized FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  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 6633)
  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..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1725..2432
                     /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(1870..1888)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2138..2157
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2445..2765
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(3657..3676)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3722..3777
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3776..3795)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4136..4152)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4136..4152)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4149..4171)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4160..4176
                     /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(4184..4190,4191..4208,4209..4214))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4229..4250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4309..4504
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4355..4490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4510..4644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4547..4566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4601..4620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4712..4729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4883..5471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4963..4982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5642..6433,6434..6502))
                     /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     6265..6284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6503..6607)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcatggtg agcaagggcg
     1741 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     1801 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1861 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     1921 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     1981 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2041 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2101 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     2161 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     2221 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2281 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2341 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2401 actccaccgg cggcatggac gagctgtaca agagtgctgg tggtggagtg caggtggaaa
     2461 ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact
     2521 acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga aacaagccct
     2581 ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga
     2641 tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg
     2701 ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac
     2761 tggaaagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     2821 gtggtcctcg aggtgcggcc gccatgaacc cctctgacca gagcccatct cctcaggact
     2881 ccacggggcc tcagctggcg accatgagac aactctcgga tgcagataag ctgcgcaagg
     2941 tgatctgcga gctcctggag acggagcgca cctacgtgaa ggatttaaac tgtcttatgg
     3001 agagatacct aaagcctctt caaaaagaaa cttttctcac ccaggatgag cttgacgtgc
     3061 tttttggaaa tttaacggaa atggtagagt ttcaagtaga attccttaaa actctagaag
     3121 atggagtgag actggtacct gatttggaaa agcttgagaa ggttgatcaa tttaagaaag
     3181 tgctgttctc tctgggggga tcattcctgt attatgctga ccgcttcaag ctctacagtg
     3241 ccttctgcgc cagccacaca aaagttccca aggtcctggt gaaagccaag acagacacgg
     3301 ctttcaaggc attcttggat gcccagaacc cgaagcagca gcactcatcc acgctggagt
     3361 cgtacctcat caagcccatc cagaggatcc tcaagtaccc acttctgctc agggagctgt
     3421 tcgccctgac cgatgcggag agcgaggagc actaccacct ggacgtggcc atcaagacca
     3481 tgaacaaggt tgccagtcac atcaatgaga tgcagaaaat ccatgaagag tttggggctg
     3541 tgtttgacca gctgattgct tgacctcgag ggcggccgca ctcctcaggt gcaggctgcc
     3601 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     3661 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     3721 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3781 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3841 ttggcaacat atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca
     3901 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3961 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     4021 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     4081 ccctcttctc ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     4141 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4201 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4261 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     4321 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4381 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4441 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4501 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4621 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     4681 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4741 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4801 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4861 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4921 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4981 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5041 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5101 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5161 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5221 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5281 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5341 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5401 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5461 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5521 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5581 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5641 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5701 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5761 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5821 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5881 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5941 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6001 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6061 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6121 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6181 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6241 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6301 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6361 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6421 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6481 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6541 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6601 ttccgcgcac atttccccga aaagtgccac ctg
//
