LOCUS       sequence-328967-        7940 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 7940)
  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 7940)
  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..7940
                     /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"
     protein_bind    1745..1778
                     /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             1821..2615
                     /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(1875..1894)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2485..2504
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2795..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2938..2971
                     /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      3007..3016
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3013..3015,3016..3018,3019..3729)
                     /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(3058..3079)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3319..3338)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3666..3687
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4964..4983)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5029..5084
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5083..5102)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5443..5459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5443..5459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5456..5478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5467..5483
                     /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(5491..5497,5498..5515,5516..5521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5536..5557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5616..5811
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5662..5797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5817..5951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6019..6036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6190..6778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6270..6289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6949..7740,7741..7809))
                     /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     7572..7591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7810..7914)
                     /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 attgattaat tcgagcgaac
     1741 gcgtgaagtt cctattctct agaaagtata ggaacttccg gatttgatct gatcaagaga
     1801 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     1861 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     1921 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     1981 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     2041 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     2101 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     2161 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     2221 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     2281 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     2341 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     2401 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     2461 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     2521 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     2581 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     2641 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     2701 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     2761 tctcatgctg gagttcttcg cccaccccat cgataacttg tttattgcag cttataatgg
     2821 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2881 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca aatccgagaa
     2941 gttcctattc tctagaaagt ataggaactt catctcgagt cgctcggtac gatttaaatt
     3001 gaattcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3061 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3121 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     3181 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc
     3241 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3301 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3361 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3421 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     3481 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     3541 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3601 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     3661 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3721 ctgtacaagt ccggactcag atctcgagct atggatgacg atatcgctgc gctggtcgtc
     3781 gacaacggct ccggcatgtg caaagccggc ttcgcgggcg acgatgctcc ccgggctgta
     3841 ttcccctcca tcgtgggccg ccctaggcac cagggtgtga tggtgggaat gggtcagaag
     3901 gactcctatg tgggtgacga ggcccagagc aagagaggta tcctgaccct gaagtacccc
     3961 attgaacatg gcattgttac caactgggac gacatggaga agatctggca ccacaccttc
     4021 tacaatgagc tgcgtgtggc ccctgaggag caccctgtgc tgctcaccga ggcccccctg
     4081 aaccctaagg ccaaccgtga aaagatgacc cagatcatgt ttgagacctt caacacccca
     4141 gccatgtacg tagccatcca ggctgtgctg tccctgtatg cctctggtcg taccacaggc
     4201 attgtgatgg actccggaga cggggtcacc cacactgtgc ccatctacga gggctatgct
     4261 ctccctcacg ccatcctgcg tctggacctg gctggccggg acctgacaga ctacctcatg
     4321 aagatcctga ccgagcgtgg ctacagcttc accaccacag ctgagaggga aatcgtgcgt
     4381 gacatcaaag agaagctgtg ctatgttgct ctagacttcg agcaggagat ggccactgcc
     4441 gcatcctctt cctccctgga gaagagctat gagctgcctg acggccaggt catcactatt
     4501 ggcaacgagc ggttccgatg ccctgaggct cttttccagc cttccttctt gggtatggaa
     4561 tcctgtggca tccatgaaac tacattcaat tccatcatga agtgtgacgt tgacatccgt
     4621 aaagacctct atgccaacac agtgctgtct ggtggtacca ccatgtaccc aggcattgct
     4681 gacaggatgc agaaggagat tactgctctg gctcctagca ccatgaagat caagatcatt
     4741 gctcctcctg agcgcaagta ctctgtgtgg atcggtggct ccatcctggc ctcactgtcc
     4801 accttccagc agatgtggat cagcaagcag gagtacgatg agtccggccc ctccatcgtg
     4861 caccgcaagt gcttctaggc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg
     4921 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     4981 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     5041 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     5101 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     5161 ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta tatgaaacag
     5221 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     5281 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     5341 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     5401 tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     5461 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5521 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5581 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
     5641 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5701 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5761 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
     5821 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5881 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5941 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6361 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     6421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6601 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7921 tccccgaaaa gtgccacctg
//
