LOCUS       sequence-340568-        7930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An actin FRET sensor. Must be expressed with ECFP-actin for FRET..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7930)
  AUTHORS   Aihara S, Fujimoto S, Sakaguchi R, Imai T
  TITLE     BMPR-2 gates activity-dependent stabilization of primary dendrites 
            during mitral cell remodeling.
  JOURNAL   Cell Rep. 2021 Jun 22;35(12):109276. doi: 
            10.1016/j.celrep.2021.109276.
  PUBMED    34161760
REFERENCE   2  (bases 1 to 7930)
  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: "Cell Rep.";
            date: "2021-06-22"; volume: "35(12):109276. doi"; pages: " 
            10.1016/j.celrep.2021.109276"
COMMENT     Sequence Label: pCAFNF-Venus-Actb
FEATURES             Location/Qualifiers
     source          1..7930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1754..1787
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             1830..2624
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1884..1903)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2494..2513
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2804..2938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2947..2980
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      3016..3025
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3022..3024,3025..3027,3028..3738)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3067..3088)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3328..3347)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3675..3696
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4973..4992)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5038..5093
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5092..5111)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5454..5470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5454..5470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5467..5489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5478..5494
                     /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(5502..5508,5509..5526,5527..5532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5547..5568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      5653..5787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5807..5941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6009..6026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6180..6768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6260..6279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6939..7730,7731..7799))
                     /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     7562..7581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7800..7904)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggc gaggcggaga ggkgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgattaatt
     1741 cgagcgaacg cgtgaagttc ctattctcta gaaagtatag gaacttccgg atttgatctg
     1801 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     1861 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     1921 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     1981 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     2041 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2101 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     2161 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2221 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2281 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2341 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2401 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     2461 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     2521 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     2581 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     2641 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     2701 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     2761 gcgcggggat ctcatgctgg agttcttcgc ccaccccatc gataacttgt ttattgcagc
     2821 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2881 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
     2941 atccgagaag ttcctattct ctagaaagta taggaacttc atctcgagtc gctcggtacg
     3001 atttaaattg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     3061 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3121 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     3181 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc
     3241 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3301 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3361 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3421 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3481 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     3541 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3601 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     3661 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3721 atggacgagc tgtacaagtc cggactcaga tctcgagcta tggatgacga tatcgctgcg
     3781 ctggtcgtcg acaacggctc cggcatgtgc aaagccggct tcgcgggcga cgatgctccc
     3841 cgggctgtat tcccctccat cgtgggccgc cctaggcacc agggtgtgat ggtgggaatg
     3901 ggtcagaagg actcctatgt gggtgacgag gcccagagca agagaggtat cctgaccctg
     3961 aagtacccca ttgaacatgg cattgttacc aactgggacg acatggagaa gatctggcac
     4021 cacaccttct acaatgagct gcgtgtggcc cctgaggagc accctgtgct gctcaccgag
     4081 gcccccctga accctaaggc caaccgtgaa aagatgaccc agatcatgtt tgagaccttc
     4141 aacaccccag ccatgtacgt agccatccag gctgtgctgt ccctgtatgc ctctggtcgt
     4201 accacaggca ttgtgatgga ctccggagac ggggtcaccc acactgtgcc catctacgag
     4261 ggctatgctc tccctcacgc catcctgcgt ctggacctgg ctggccggga cctgacagac
     4321 tacctcatga agatcctgac cgagcgtggc tacagcttca ccaccacagc tgagagggaa
     4381 atcgtgcgtg acatcaaaga gaagctgtgc tatgttgctc tagacttcga gcaggagatg
     4441 gccactgccg catcctcttc ctccctggag aagagctatg agctgcctga cggccaggtc
     4501 atcactattg gcaacgagcg gttccgatgc cctgaggctc ttttccagcc ttccttcttg
     4561 ggtatggaat cctgtggcat ccatgaaact acattcaatt ccatcatgaa gtgtgacgtt
     4621 gacatccgta aagacctcta tgccaacaca gtgctgtctg gtggtaccac catgtaccca
     4681 ggcattgctg acaggatgca gaaggagatt actgctctgg ctcctagcac catgaagatc
     4741 aagatcattg ctcctcctga gcgcaagtac tctgtgtgga tcggtggctc catcctggcc
     4801 tcactgtcca ccttccagca gatgtggatc agcaagcagg agtacgatga gtccggcccc
     4861 tccatcgtgc accgcaagtg cttctaggcg gccgcactcc tcaggtgcag gctgcctatc
     4921 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     4981 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     5041 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     5101 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     5161 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     5221 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     5281 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     5341 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     5401 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     5461 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5521 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5581 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     5641 cagcaaccat agtcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5701 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5761 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     5821 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5881 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     5941 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6001 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6061 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6121 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6181 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6241 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6301 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6361 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6421 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6481 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6541 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6601 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6661 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6721 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6781 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6841 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6901 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6961 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7021 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7081 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7141 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7201 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7261 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7321 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7381 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7441 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7501 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7561 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7621 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7681 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7741 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7801 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7861 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7921 gtgccacctg
//
