LOCUS       sequence-165638-        6373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FLIM acceptor for mEGFP Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  AUTHORS   Hedrick NG, Harward SC, Hall CE, Murakoshi H, McNamara JO, Yasuda R
  TITLE     Rho GTPase complementation underlies BDNF-dependent homo- and 
            heterosynaptic plasticity.
  JOURNAL   Nature. 2016 Sep 28;538(7623):104-108. doi: 10.1038/nature19784.
  PUBMED    27680697
REFERENCE   2  (bases 1 to 6373)
  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"; journalName: 
            "Nature"; date: "2016-09-28- 28"; volume: "538"; issue: "7623"; 
            pages: "104-108"
COMMENT     Sequence Label: mCherry-PAK2 binding domain
FEATURES             Location/Qualifiers
     source          1..6373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1748..1757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1754..2461
                     /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"
     CDS             2660..3370
                     /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(3397..3416)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3462..3517
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3516..3535)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3876..3892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3876..3892)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3889..3911)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3900..3916
                     /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(3924..3930,3931..3948,3949..3954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3969..3990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4049..4244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4095..4230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4250..4384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4287..4306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4341..4360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4452..4469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4623..5211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4703..4722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5382..6173,6174..6242))
                     /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     6005..6024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6243..6347)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cgggggcgag
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaagaatt cgctagcgct
     1741 accggtcgcc accatggtga gcaagggcga ggaggacaac atggccatca tcaaggagtt
     1801 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     1861 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1921 cggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     1981 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2041 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2101 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2161 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     2221 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     2281 ccactacgac gccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     2341 cgcctacaac gtcaacatca agctggacat cacctcccac aacgaggact acaccatcgt
     2401 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     2461 gtccggactc agatctcgag gaggaatgaa caaaaagaaa gagaaagagt gcccagagat
     2521 ttctccccct gcagattttg aacacacaat tcatgtcggt tttgatgctg tcacagggga
     2581 gtttacggga atgccagagc agtgggcccg cttgcttcag acatcaaata tcactaagtt
     2641 ggagcagaag ggatccggaa tggtgagcaa gggcgaggag gacaacatgg ccatcatcaa
     2701 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     2761 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     2821 caagggcggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     2881 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     2941 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     3001 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     3061 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     3121 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     3181 cggcggccac tacgacgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     3241 gcccggcgcc tacaacgtca acatcaagct ggacatcacc tcccacaacg aggactacac
     3301 catcgtggaa cagtacgagc gcgccgaggg ccgccactcc accggcggca tggacgagct
     3361 gtacaagtaa gcggccgcga ctctagagtc gacggatctt tttccctctg ccaaaaatta
     3421 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     3481 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3541 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgccatatg
     3601 ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa cagccccctg
     3661 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt ttttatattt
     3721 tgtttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     3781 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatgaagat
     3841 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3901 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3961 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4021 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     4081 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4141 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4201 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     4261 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4321 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4381 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4801 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5041 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6361 aaagtgccac ctg
//
