LOCUS       sequence-219989-        9225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the PH 
            domain from Human pleckstrin homology domain containing A3 
            (PLEKHA3). Binds PtdIns4P.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9225)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 9225)
  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: "Nat 
            Commun."; date: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: actc1b-mKate2-FAPP1
FEATURES             Location/Qualifiers
     source          1..9225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      26..35
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             32..727
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     protein_bind    complement(729..753)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1049..1069)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(1077..1093)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1109..1243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1133..1152)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1187..1206
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1682..1869,1870..1881)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1915..1931)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1915..1931)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1968..1988)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1968..1986)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2007..2023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2020..2042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2031..2047
                     /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(2055..2061,2062..2079,2080..2085))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2100..2121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2238..2255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2409..2997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2489..2508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3168..3959,3960..4028))
                     /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     3791..3810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4029..4133)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4296..4317)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4666..4677,4678..4840)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5213..5229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5249..5269
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 gcaagtttgt acaaaaaagc aggctgccac catggtgagc gagctgatta aggagaacat
       61 gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga
      121 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg
      181 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt
      241 catcaaccac acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac
      301 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
      361 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa
      421 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc
      481 cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca
      541 cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat
      601 gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac
      661 ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg
      721 gcacagagac ccagctttct tgtacaaagt ggctgagggg gtgttgtaca agtggaccaa
      781 ctatctcaca ggctggcagc ctcgttggtt tgttttagat aatggaatct tatcctacta
      841 tgattcacaa gatgatgttt gcaaagggag caaaggaagc ataaagatgg cagtttgtga
      901 aattaaagtt cattcagcag acaacacaag aatggaatta atcattcctg gagagcagca
      961 tttctacatg aaggcagtga atgcagctga aagacagagg tggctggtcg ctctggggag
     1021 ctccaaagca tgtttgactg atacatgaca actttattat acatagttga taattcactg
     1081 gccgtcgttt tacggtacca tcgatgatga tccagacatg ataagataca ttgatgagtt
     1141 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1201 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1261 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     1321 ctacaaatgt ggtatggctg attatgatcc tctagatcag atctgcgaag atacggccac
     1381 gggtgctctt gatcctgtgg ctgattttgg actgtgctgc tcgcagctgc tgatgaatca
     1441 catacttcct ccattttctt ccactgattg actgttataa tttccctaat ttccaggtca
     1501 aggtgctgtg cattgtggta atagatgtga catgacgtca cttccaaagg accaatgaac
     1561 atgtctgacc aatttcatat aatgtgaaaa cgattttcat aggcagaata aataacattt
     1621 aaattaaact gggcatcagc gcaattcaat tggtttggta atagcaaggg aaaatagaat
     1681 gaagtgatct ccaaaaaata agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa
     1741 gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa aagtattgat ttttaattgt
     1801 actcaagtaa agtaaaaatc cccaaaaata atacttaagt acagtaatca agtaaaatta
     1861 ctcaagtact ttacacctct ggttcttgac cccctacctt cagcaagccc agcagatcca
     1921 ctagttctag agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg
     1981 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2041 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2101 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2161 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2221 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2281 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2341 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2401 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2461 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2521 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2581 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2641 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2701 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2761 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2821 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2881 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2941 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3001 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggc
     3061 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3121 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3181 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3241 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3301 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3361 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3421 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3481 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3541 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3601 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3661 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3721 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3781 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3841 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3901 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3961 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4021 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4081 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4141 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4201 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4261 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4321 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4381 gctggctaag aactcatcag cctccccggt ccatctaccc acgtaccaat gcaccaattg
     4441 gccacaatga cggctactac atggtgccat tccttcctct ttataggaat ggagactacc
     4501 tcctgtccaa caatgctctt ggatacgagt acgcctacct gttggaccca ggtcattgca
     4561 caacaccaga aatgccctct gatctgcaaa agacgtgaat atctgttcag acacccatat
     4621 ccactctgtt ccacacaggt cagaggtttg tccaggagtt cttgacagag gtgtaaaaag
     4681 tactcaaaaa ttttactcaa gtgaaagtac aagtacttag ggaaaatttt actcaattaa
     4741 aagtaaaagt atctggctag aatcttactt gagtaaaagt aaaaaagtac tccattaaaa
     4801 ttgtacttga gtattaagga agtaaaagta aaagcaagaa agaaaactag agattcttgt
     4861 ttaagctttt aatctcaaaa aacattaaat gaaatgcata caaggtttta tcctgcttta
     4921 gaactgtttg tatttaatta tcaaactata agacagacaa tctaatgcca gtacacgcta
     4981 ctcaaagttg taaaacctca gatttaactt cagtagaagc tgattctcaa aattgttagt
     5041 gtcaagccta gctcttttgg ggctgaaaag caatcctgca gtgctgaaaa gcctctcaca
     5101 ggcagccgat gcgggaagag gtgtattagt cttgatagag aggctgcaaa tagcaggaaa
     5161 cgtgagcaga gactccctgg tgtctgaaac acaggccaga tgggccctcg agcaggaaac
     5221 agctatgacc atgattacgc caagctatca actttgtata gaaaagttgg cacgcgcaga
     5281 ttccgtgtgg gcctggtaat atcttaccat tttaagctat tttaagatta gggaaaaaaa
     5341 ttaaataaaa ctactttaaa attactttcc ttaaaaagca acatatttat ttagtttttg
     5401 tttaactggc tcaagattgt aatattttac ttttacatgt ttttttaagt acctaaatgt
     5461 gttcaaaagt aaagtatgtt attgtttttt ttttttaatg ttagctaagc tttaaattgt
     5521 aatatattac ttttaaaagt gactttttaa aaatgtaaat aaaaaagtaa ctataaatgt
     5581 gtaatgcatg ttatttctgt gggtttttaa ttttcatctc aactgggctt acaattttaa
     5641 cataataaaa atcaaatcaa atcaaatcag gcccttattt gtcacataca ctcttgtata
     5701 ctgaaataaa ctaaatgttg agaaaaattt aaatacacca aaataacgca tacctttttt
     5761 gtaattacct aagtaaaaca tttagttcct tttttaagtg actcaagatt gtaatctatt
     5821 aattgtaaaa gataaaacta gtgatcaatc tgacgtgctt ctatgcagcc tgtgagatta
     5881 attgtgatgt ttaattatta aacaatttaa atggatcaaa catttcacat gtgcttattg
     5941 tgtccaacaa aaccagcaga agcacttaaa aatattgcat gacaacatga gtttttaatg
     6001 taaattatat atacttccta aacgaagtta atcatatttc taaaataata atgtcattga
     6061 aatatttaaa taaatgataa gctttagttg attcaacata aaattgaaag caataaggac
     6121 tttttttttt tacagcggtg aagcaaaagc atgtaataat catataatat aataaataaa
     6181 taaataaata atggctcttg gtgcaaatat gaacacttta aataaatata gaataaatat
     6241 gatcaataag tcataacgat gtgttttatg ttgattataa tttaaacatg tttctaacgc
     6301 aagttttaaa aactgtttta aagtattatt ttaagttata agatcaattt gactcaacat
     6361 tttgtagtga aaatatgttg tgtaaaatta atataatata cactataaaa atgcagggtt
     6421 ccacacaata cattaatgtt gtcccaacac aaatcgatta agttaacaaa tatatgtgga
     6481 ttgaacataa aaaattaagt tgtcccaatg aaatctcaag aattgtgttg tttcagctca
     6541 ttttaaataa gtagttttaa caagtaaaaa ataacatatt tttgagtgtg tatatatata
     6601 tattacaaaa tataataata taaaatgtaa aagatataaa ccatgctaca tatatttaag
     6661 agctattttg cacattactt ttttgcaagc cattttagtg ttttttttac aaatatatat
     6721 tagctaaata tacaatatct taaatataca attaatcaga catgcaatta atgcatttaa
     6781 cctacataaa aatacatgat caacatatgt tttatgctga ttacaactaa atcaaacgta
     6841 atgaaattaa tttataaaag cgatttaaat atgtcaatat tttaagttaa aatcacttga
     6901 aacctactat aaatgaactt aaattaaaaa aatctccttt aattccaatg caacagtgtt
     6961 gttttttcag ctgaggaccc ccggtgcaga tggttggccc ccggcccgct attctaagga
     7021 agcctttctt aattttaggg cggcgctgca tgcgctcaga tcagattcac gggtggagag
     7081 ggttattttc tcttttttat tttttcttgc ctgctcatat ttgggcctgt gtgaacccca
     7141 ggctccgcct cggcccccag gagccgctct tatttaaagg ggatcctcgc cttcagtctt
     7201 cgtcccagtg gaagcatcag tcttgtgata acaacttggt gagtcctaca gaccagcgct
     7261 cgactctagg ctttaaatgc attgatttct ctggattagc taccgggaga gttttaactt
     7321 tgcttattag tcttgtgctc gaactgctag gtttaaagag tgttttctct aatgcatgca
     7381 tgaagcgaca gctgaaaacc agaaatagag cgtttgtcct cctgttaaac tctagccttt
     7441 ctttggcatt tgtcagtccg agtcttcagc tgccggcagg agattgggtt tcagtaacac
     7501 ttgctgagca caaactacac gtgctttctt cagatttaat tcattattaa tcattctaga
     7561 tttaaatgat gttcttactc attgctgatt attttagcct tgcggagcag attaaatgag
     7621 cacgtgtttc attaagattg atttaaatat aattaatatt tcaccactgt agttttaaaa
     7681 ttgccttcta agtcatggtt gatttttaat aggtatcggc tctaattaga ttactatttt
     7741 ggtttttaaa tgttatttct atttcatttg taggctcgtt tttaatgtga ttttattaga
     7801 ttctagtttt aaaatcatct cttagcctta atgagcagac tgaatgaaca tgtttcatta
     7861 agattaactt cattataatg aattatttaa tcattctagt gggttataaa tcagctctat
     7921 ttaaattaat tgtcgtgttt atttctattt aatttttgat tattttttca gggtcgtttt
     7981 tatgtgattt tttcccgttt ctaattttaa actaattgtt tgatttttat atgattcgac
     8041 tctttttaaa tgaattgttt tgcttttttt attgagcata tatatattat acaatttcaa
     8101 tcaacaattg ttgtgctttt taaatgttgt ttctatttca tttctagggt cgattttatt
     8161 cgattctagt tttaaaatcg tgatttttaa cttctaacct tactgagcag actgaatgaa
     8221 catgtttcat caagattaac ttcattatag ttaattattg aatcgtgcta gtggtttata
     8281 aatcagctct atttaaatta atcatcatca ttatttctat ttaattttta ttatttcttt
     8341 aggattattt ttatgtgatt tttttcacgt tctagtttta aactcaatgc tcattactgc
     8401 tcacccttgc tgagcagtct ttcgttaaga ttaaattaat taataaccat tctagtttta
     8461 aaacagtgtc ttaaggcata gctgatattt tacaagttta ctatttaata tatctccttt
     8521 ctttttttag ggtcgatttc atctgatttg attaattttt attatcttca ctaagctttt
     8581 tgttctaggt tttattctca gtattattac tgttatattt gaactgaatt tctcctcata
     8641 gtttattcat tttatttatt tattttttgt ttattcgcta ttttgtattt tagtttttaa
     8701 ttagaaattt tattagaatc gttttgaact agtttttatg cactctaaca ttaatattaa
     8761 atcataacta ttatttttat tataatttat ttgtgtttat ctttttgtgt agctttcatg
     8821 tgtttttatt ttagttttat tatgtttgca ttgaattttt tggactagtt tatataacga
     8881 tatacagaaa agccatgaca acaaatgttt ttataatacc tattaagtat aagctaatca
     8941 gatttccact attctgttca aagttaacat tattagccat tctgatggat catttgacat
     9001 ttgtcatatg tctttatgaa cactgctctt tcagatgcca cttttaaagc tgttttttac
     9061 agtaaatgaa cgtttattat tatgacatta tttcctaagg cctctagagt acagcaggat
     9121 gtggcgtgtg cattaccgtg taattatgct gactgtaaat gtgcatttat agactgtcag
     9181 catgtctgac tgtgcgctgt tgtgattggt cctgcacaga ccaag
//
