LOCUS       sequence-224841-        9446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to mouse 2x PH 
            domain from HGF regulated tyrosine kinase substrate (Hgs). Binds 
            PtdIns3P.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9446)
  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 9446)
  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-mKate2-2xFYVEhgs
FEATURES             Location/Qualifiers
     source          1..9446
                     /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(1270..1290)
                     /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(1298..1314)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1330..1464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1354..1373)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1408..1427
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1903..2090,2091..2102)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2136..2152)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2136..2152)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2189..2209)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2189..2207)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2228..2244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2241..2263)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2252..2268
                     /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(2276..2282,2283..2300,2301..2306))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2321..2342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2459..2476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2630..3218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2710..2729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3389..4180,4181..4249))
                     /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     4012..4031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4250..4354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4517..4538)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4887..4898,4899..5061)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5434..5450
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5470..5490
                     /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 ggctctcgag ctcaagcttc gaattgaatt
      781 cgaaagtgat gccatgttcg ctgctgaaag agcccctgac tgggtggatg ctgaggaatg
      841 ccatcggtgc agagtacagt ttggggtggt gacccgcaag catcactgcc gagcatgtgg
      901 gcagatcttc tgtggcaagt gctcctccaa gtactccacc atccccaagt tcggcattga
      961 gaaggaggtg cgcgtgtgtg agccctgcta tgagcagctg aacaagaagg cacaaggaca
     1021 aggatccgaa agtgatgcca tgttcgctgc tgaaagagcc cctgactggg tggatgctga
     1081 ggaatgccat cggtgcagag tacagtttgg ggtggtgacc cgcaagcatc actgccgagc
     1141 atgtgggcag atcttctgtg gcaagtgctc ctccaagtac tccaccatcc ccaagttcgg
     1201 cattgagaag gaggtgcgcg tgtgtgagcc ctgctatgag cagctgaaca agaaggcata
     1261 agtcgacggc aactttatta tacatagttg ataattcact ggccgtcgtt ttacggtacc
     1321 atcgatgatg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1381 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1441 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1501 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     1561 gattatgatc ctctagatca gatctgcgaa gatacggcca cgggtgctct tgatcctgtg
     1621 gctgattttg gactgtgctg ctcgcagctg ctgatgaatc acatacttcc tccattttct
     1681 tccactgatt gactgttata atttccctaa tttccaggtc aaggtgctgt gcattgtggt
     1741 aatagatgtg acatgacgtc acttccaaag gaccaatgaa catgtctgac caatttcata
     1801 taatgtgaaa acgattttca taggcagaat aaataacatt taaattaaac tgggcatcag
     1861 cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat
     1921 aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa
     1981 aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat
     2041 ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc
     2101 tggttcttga ccccctacct tcagcaagcc cagcagatcc actagttcta gagcggccgc
     2161 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     2221 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2281 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2341 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2401 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2461 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2521 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2581 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2641 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2701 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2761 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2821 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2881 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2941 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3001 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3061 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3121 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3181 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3241 cggggtctga cgctcagtgg aacgaaaact cacgttaagg cattttggtc atgagattat
     3301 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3361 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3421 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3481 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3541 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3601 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3661 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3721 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3781 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3841 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3901 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3961 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4021 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4081 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4141 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4201 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4261 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4321 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     4381 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4441 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4501 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4561 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca cgctggctaa gaactcatca
     4621 gcctccccgg tccatctacc cacgtaccaa tgcaccaatt ggccacaatg acggctacta
     4681 catggtgcca ttccttcctc tttataggaa tggagactac ctcctgtcca acaatgctct
     4741 tggatacgag tacgcctacc tgttggaccc aggtcattgc acaacaccag aaatgccctc
     4801 tgatctgcaa aagacgtgaa tatctgttca gacacccata tccactctgt tccacacagg
     4861 tcagaggttt gtccaggagt tcttgacaga ggtgtaaaaa gtactcaaaa attttactca
     4921 agtgaaagta caagtactta gggaaaattt tactcaatta aaagtaaaag tatctggcta
     4981 gaatcttact tgagtaaaag taaaaaagta ctccattaaa attgtacttg agtattaagg
     5041 aagtaaaagt aaaagcaaga aagaaaacta gagattcttg tttaagcttt taatctcaaa
     5101 aaacattaaa tgaaatgcat acaaggtttt atcctgcttt agaactgttt gtatttaatt
     5161 atcaaactat aagacagaca atctaatgcc agtacacgct actcaaagtt gtaaaacctc
     5221 agatttaact tcagtagaag ctgattctca aaattgttag tgtcaagcct agctcttttg
     5281 gggctgaaaa gcaatcctgc agtgctgaaa agcctctcac aggcagccga tgcgggaaga
     5341 ggtgtattag tcttgataga gaggctgcaa atagcaggaa acgtgagcag agactccctg
     5401 gtgtctgaaa cacaggccag atgggccctc gagcaggaaa cagctatgac catgattacg
     5461 ccaagctatc aactttgtat agaaaagttg gcacgcgcag attccgtgtg ggcctggtaa
     5521 tatcttacca ttttaagcta ttttaagatt agggaaaaaa attaaataaa actactttaa
     5581 aattactttc cttaaaaagc aacatattta tttagttttt gtttaactgg ctcaagattg
     5641 taatatttta cttttacatg tttttttaag tacctaaatg tgttcaaaag taaagtatgt
     5701 tattgttttt tttttttaat gttagctaag ctttaaattg taatatatta cttttaaaag
     5761 tgacttttta aaaatgtaaa taaaaaagta actataaatg tgtaatgcat gttatttctg
     5821 tgggttttta attttcatct caactgggct tacaatttta acataataaa aatcaaatca
     5881 aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa actaaatgtt
     5941 gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc taagtaaaac
     6001 atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa agataaaact
     6061 agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg tttaattatt
     6121 aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca aaaccagcag
     6181 aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata tatacttcct
     6241 aaacgaagtt aatcatattt ctaaaataat aatgtcattg aaatatttaa ataaatgata
     6301 agctttagtt gattcaacat aaaattgaaa gcaataagga cttttttttt ttacagcggt
     6361 gaagcaaaag catgtaataa tcatataata taataaataa ataaataaat aatggctctt
     6421 ggtgcaaata tgaacacttt aaataaatat agaataaata tgatcaataa gtcataacga
     6481 tgtgttttat gttgattata atttaaacat gtttctaacg caagttttaa aaactgtttt
     6541 aaagtattat tttaagttat aagatcaatt tgactcaaca ttttgtagtg aaaatatgtt
     6601 gtgtaaaatt aatataatat acactataaa aatgcagggt tccacacaat acattaatgt
     6661 tgtcccaaca caaatcgatt aagttaacaa atatatgtgg attgaacata aaaaattaag
     6721 ttgtcccaat gaaatctcaa gaattgtgtt gtttcagctc attttaaata agtagtttta
     6781 acaagtaaaa aataacatat ttttgagtgt gtatatatat atattacaaa atataataat
     6841 ataaaatgta aaagatataa accatgctac atatatttaa gagctatttt gcacattact
     6901 tttttgcaag ccattttagt gtttttttta caaatatata ttagctaaat atacaatatc
     6961 ttaaatatac aattaatcag acatgcaatt aatgcattta acctacataa aaatacatga
     7021 tcaacatatg ttttatgctg attacaacta aatcaaacgt aatgaaatta atttataaaa
     7081 gcgatttaaa tatgtcaata ttttaagtta aaatcacttg aaacctacta taaatgaact
     7141 taaattaaaa aaatctcctt taattccaat gcaacagtgt tgttttttca gctgaggacc
     7201 cccggtgcag atggttggcc cccggcccgc tattctaagg aagcctttct taattttagg
     7261 gcggcgctgc atgcgctcag atcagattca cgggtggaga gggttatttt ctctttttta
     7321 ttttttcttg cctgctcata tttgggcctg tgtgaacccc aggctccgcc tcggccccca
     7381 ggagccgctc ttatttaaag gggatcctcg ccttcagtct tcgtcccagt ggaagcatca
     7441 gtcttgtgat aacaacttgg tgagtcctac agaccagcgc tcgactctag gctttaaatg
     7501 cattgatttc tctggattag ctaccgggag agttttaact ttgcttatta gtcttgtgct
     7561 cgaactgcta ggtttaaaga gtgttttctc taatgcatgc atgaagcgac agctgaaaac
     7621 cagaaataga gcgtttgtcc tcctgttaaa ctctagcctt tctttggcat ttgtcagtcc
     7681 gagtcttcag ctgccggcag gagattgggt ttcagtaaca cttgctgagc acaaactaca
     7741 cgtgctttct tcagatttaa ttcattatta atcattctag atttaaatga tgttcttact
     7801 cattgctgat tattttagcc ttgcggagca gattaaatga gcacgtgttt cattaagatt
     7861 gatttaaata taattaatat ttcaccactg tagttttaaa attgccttct aagtcatggt
     7921 tgatttttaa taggtatcgg ctctaattag attactattt tggtttttaa atgttatttc
     7981 tatttcattt gtaggctcgt ttttaatgtg attttattag attctagttt taaaatcatc
     8041 tcttagcctt aatgagcaga ctgaatgaac atgtttcatt aagattaact tcattataat
     8101 gaattattta atcattctag tgggttataa atcagctcta tttaaattaa ttgtcgtgtt
     8161 tatttctatt taatttttga ttattttttc agggtcgttt ttatgtgatt ttttcccgtt
     8221 tctaatttta aactaattgt ttgattttta tatgattcga ctctttttaa atgaattgtt
     8281 ttgctttttt tattgagcat atatatatta tacaatttca atcaacaatt gttgtgcttt
     8341 ttaaatgttg tttctatttc atttctaggg tcgattttat tcgattctag ttttaaaatc
     8401 gtgattttta acttctaacc ttactgagca gactgaatga acatgtttca tcaagattaa
     8461 cttcattata gttaattatt gaatcgtgct agtggtttat aaatcagctc tatttaaatt
     8521 aatcatcatc attatttcta tttaattttt attatttctt taggattatt tttatgtgat
     8581 ttttttcacg ttctagtttt aaactcaatg ctcattactg ctcacccttg ctgagcagtc
     8641 tttcgttaag attaaattaa ttaataacca ttctagtttt aaaacagtgt cttaaggcat
     8701 agctgatatt ttacaagttt actatttaat atatctcctt tcttttttta gggtcgattt
     8761 catctgattt gattaatttt tattatcttc actaagcttt ttgttctagg ttttattctc
     8821 agtattatta ctgttatatt tgaactgaat ttctcctcat agtttattca ttttatttat
     8881 ttattttttg tttattcgct attttgtatt ttagttttta attagaaatt ttattagaat
     8941 cgttttgaac tagtttttat gcactctaac attaatatta aatcataact attattttta
     9001 ttataattta tttgtgttta tctttttgtg tagctttcat gtgtttttat tttagtttta
     9061 ttatgtttgc attgaatttt ttggactagt ttatataacg atatacagaa aagccatgac
     9121 aacaaatgtt tttataatac ctattaagta taagctaatc agatttccac tattctgttc
     9181 aaagttaaca ttattagcca ttctgatgga tcatttgaca tttgtcatat gtctttatga
     9241 acactgctct ttcagatgcc acttttaaag ctgtttttta cagtaaatga acgtttatta
     9301 ttatgacatt atttcctaag gcctctagag tacagcagga tgtggcgtgt gcattaccgt
     9361 gtaattatgc tgactgtaaa tgtgcattta tagactgtca gcatgtctga ctgtgcgctg
     9421 ttgtgattgg tcctgcacag accaag
//
