LOCUS       sequence-219926-        9185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the actin 
            binding lifeact sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9185)
  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 9185)
  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-Lifeact-mKate2
FEATURES             Location/Qualifiers
     source          1..9185
                     /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..82
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     protein_bind    complement(84..108)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             110..805
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     polyA_signal    927..1008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1009..1029)
                     /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(1037..1053)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1069..1203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1093..1112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(1642..1829,1830..1841)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1875..1891)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1875..1891)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1928..1948)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1928..1946)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1967..1983)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1980..2002)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1991..2007
                     /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(2015..2021,2022..2039,2040..2045))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2060..2081
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2198..2215)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2369..2957)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2449..2468)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3128..3919,3920..3988))
                     /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     3751..3770
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3989..4093)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4256..4277)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4626..4637,4638..4800)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5173..5189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5209..5229
                     /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 catgggcgtg gccgacctga tcaagaagtt
       61 cgagagcatc agcaaggagg agaacccagc tttcttgtac aaagtggctg tgagcgagct
      121 gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt
      181 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa
      241 ggcggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta
      301 cggcagcaaa accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt
      361 ccccgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc
      421 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt
      481 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggcctccac
      541 cgagaccctg taccccgctg acggcggcct ggaaggcaga gccgacatgg ccctgaagct
      601 cgtgggcggg ggccacctga tctgcaactt gaagaccaca tacagatcca agaaacccgc
      661 taagaacctc aagatgcccg gcgtctacta tgtggacaga agactggaaa gaatcaagga
      721 ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct
      781 ccctagcaaa ctggggcaca gatgagcggc cgcgactcta gatcataatc agccatacca
      841 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
      901 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
      961 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcca actttattat
     1021 acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga tccagacatg
     1081 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1141 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1201 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1261 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc tctagatcag
     1321 atctgcgaag atacggccac gggtgctctt gatcctgtgg ctgattttgg actgtgctgc
     1381 tcgcagctgc tgatgaatca catacttcct ccattttctt ccactgattg actgttataa
     1441 tttccctaat ttccaggtca aggtgctgtg cattgtggta atagatgtga catgacgtca
     1501 cttccaaagg accaatgaac atgtctgacc aatttcatat aatgtgaaaa cgattttcat
     1561 aggcagaata aataacattt aaattaaact gggcatcagc gcaattcaat tggtttggta
     1621 atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt gactgtaaat
     1681 aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa
     1741 aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata atacttaagt
     1801 acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac cccctacctt
     1861 cagcaagccc agcagatcca ctagttctag agcggccgcc accgcggtgg agctccagct
     1921 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     1981 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2041 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2101 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2161 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2221 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2281 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2341 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2401 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2461 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2521 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2581 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2641 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2701 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2761 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2821 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2881 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2941 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3001 acgaaaactc acgttaaggc attttggtca tgagattatc aaaaaggatc ttcacctaga
     3061 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3121 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3181 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3241 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3301 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3361 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3421 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3481 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3541 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3601 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3661 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3721 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3781 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3841 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3901 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3961 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4021 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4081 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     4141 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4201 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4261 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4321 ctatcagggc gatggcccac gctggctaag aactcatcag cctccccggt ccatctaccc
     4381 acgtaccaat gcaccaattg gccacaatga cggctactac atggtgccat tccttcctct
     4441 ttataggaat ggagactacc tcctgtccaa caatgctctt ggatacgagt acgcctacct
     4501 gttggaccca ggtcattgca caacaccaga aatgccctct gatctgcaaa agacgtgaat
     4561 atctgttcag acacccatat ccactctgtt ccacacaggt cagaggtttg tccaggagtt
     4621 cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac aagtacttag
     4681 ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt gagtaaaagt
     4741 aaaaaagtac tccattaaaa ttgtacttga gtattaagga agtaaaagta aaagcaagaa
     4801 agaaaactag agattcttgt ttaagctttt aatctcaaaa aacattaaat gaaatgcata
     4861 caaggtttta tcctgcttta gaactgtttg tatttaatta tcaaactata agacagacaa
     4921 tctaatgcca gtacacgcta ctcaaagttg taaaacctca gatttaactt cagtagaagc
     4981 tgattctcaa aattgttagt gtcaagccta gctcttttgg ggctgaaaag caatcctgca
     5041 gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag gtgtattagt cttgatagag
     5101 aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg tgtctgaaac acaggccaga
     5161 tgggccctcg agcaggaaac agctatgacc atgattacgc caagctatca actttgtata
     5221 gaaaagttgg cacgcgcaga ttccgtgtgg gcctggtaat atcttaccat tttaagctat
     5281 tttaagatta gggaaaaaaa ttaaataaaa ctactttaaa attactttcc ttaaaaagca
     5341 acatatttat ttagtttttg tttaactggc tcaagattgt aatattttac ttttacatgt
     5401 ttttttaagt acctaaatgt gttcaaaagt aaagtatgtt attgtttttt ttttttaatg
     5461 ttagctaagc tttaaattgt aatatattac ttttaaaagt gactttttaa aaatgtaaat
     5521 aaaaaagtaa ctataaatgt gtaatgcatg ttatttctgt gggtttttaa ttttcatctc
     5581 aactgggctt acaattttaa cataataaaa atcaaatcaa atcaaatcag gcccttattt
     5641 gtcacataca ctcttgtata ctgaaataaa ctaaatgttg agaaaaattt aaatacacca
     5701 aaataacgca tacctttttt gtaattacct aagtaaaaca tttagttcct tttttaagtg
     5761 actcaagatt gtaatctatt aattgtaaaa gataaaacta gtgatcaatc tgacgtgctt
     5821 ctatgcagcc tgtgagatta attgtgatgt ttaattatta aacaatttaa atggatcaaa
     5881 catttcacat gtgcttattg tgtccaacaa aaccagcaga agcacttaaa aatattgcat
     5941 gacaacatga gtttttaatg taaattatat atacttccta aacgaagtta atcatatttc
     6001 taaaataata atgtcattga aatatttaaa taaatgataa gctttagttg attcaacata
     6061 aaattgaaag caataaggac tttttttttt tacagcggtg aagcaaaagc atgtaataat
     6121 catataatat aataaataaa taaataaata atggctcttg gtgcaaatat gaacacttta
     6181 aataaatata gaataaatat gatcaataag tcataacgat gtgttttatg ttgattataa
     6241 tttaaacatg tttctaacgc aagttttaaa aactgtttta aagtattatt ttaagttata
     6301 agatcaattt gactcaacat tttgtagtga aaatatgttg tgtaaaatta atataatata
     6361 cactataaaa atgcagggtt ccacacaata cattaatgtt gtcccaacac aaatcgatta
     6421 agttaacaaa tatatgtgga ttgaacataa aaaattaagt tgtcccaatg aaatctcaag
     6481 aattgtgttg tttcagctca ttttaaataa gtagttttaa caagtaaaaa ataacatatt
     6541 tttgagtgtg tatatatata tattacaaaa tataataata taaaatgtaa aagatataaa
     6601 ccatgctaca tatatttaag agctattttg cacattactt ttttgcaagc cattttagtg
     6661 ttttttttac aaatatatat tagctaaata tacaatatct taaatataca attaatcaga
     6721 catgcaatta atgcatttaa cctacataaa aatacatgat caacatatgt tttatgctga
     6781 ttacaactaa atcaaacgta atgaaattaa tttataaaag cgatttaaat atgtcaatat
     6841 tttaagttaa aatcacttga aacctactat aaatgaactt aaattaaaaa aatctccttt
     6901 aattccaatg caacagtgtt gttttttcag ctgaggaccc ccggtgcaga tggttggccc
     6961 ccggcccgct attctaagga agcctttctt aattttaggg cggcgctgca tgcgctcaga
     7021 tcagattcac gggtggagag ggttattttc tcttttttat tttttcttgc ctgctcatat
     7081 ttgggcctgt gtgaacccca ggctccgcct cggcccccag gagccgctct tatttaaagg
     7141 ggatcctcgc cttcagtctt cgtcccagtg gaagcatcag tcttgtgata acaacttggt
     7201 gagtcctaca gaccagcgct cgactctagg ctttaaatgc attgatttct ctggattagc
     7261 taccgggaga gttttaactt tgcttattag tcttgtgctc gaactgctag gtttaaagag
     7321 tgttttctct aatgcatgca tgaagcgaca gctgaaaacc agaaatagag cgtttgtcct
     7381 cctgttaaac tctagccttt ctttggcatt tgtcagtccg agtcttcagc tgccggcagg
     7441 agattgggtt tcagtaacac ttgctgagca caaactacac gtgctttctt cagatttaat
     7501 tcattattaa tcattctaga tttaaatgat gttcttactc attgctgatt attttagcct
     7561 tgcggagcag attaaatgag cacgtgtttc attaagattg atttaaatat aattaatatt
     7621 tcaccactgt agttttaaaa ttgccttcta agtcatggtt gatttttaat aggtatcggc
     7681 tctaattaga ttactatttt ggtttttaaa tgttatttct atttcatttg taggctcgtt
     7741 tttaatgtga ttttattaga ttctagtttt aaaatcatct cttagcctta atgagcagac
     7801 tgaatgaaca tgtttcatta agattaactt cattataatg aattatttaa tcattctagt
     7861 gggttataaa tcagctctat ttaaattaat tgtcgtgttt atttctattt aatttttgat
     7921 tattttttca gggtcgtttt tatgtgattt tttcccgttt ctaattttaa actaattgtt
     7981 tgatttttat atgattcgac tctttttaaa tgaattgttt tgcttttttt attgagcata
     8041 tatatattat acaatttcaa tcaacaattg ttgtgctttt taaatgttgt ttctatttca
     8101 tttctagggt cgattttatt cgattctagt tttaaaatcg tgatttttaa cttctaacct
     8161 tactgagcag actgaatgaa catgtttcat caagattaac ttcattatag ttaattattg
     8221 aatcgtgcta gtggtttata aatcagctct atttaaatta atcatcatca ttatttctat
     8281 ttaattttta ttatttcttt aggattattt ttatgtgatt tttttcacgt tctagtttta
     8341 aactcaatgc tcattactgc tcacccttgc tgagcagtct ttcgttaaga ttaaattaat
     8401 taataaccat tctagtttta aaacagtgtc ttaaggcata gctgatattt tacaagttta
     8461 ctatttaata tatctccttt ctttttttag ggtcgatttc atctgatttg attaattttt
     8521 attatcttca ctaagctttt tgttctaggt tttattctca gtattattac tgttatattt
     8581 gaactgaatt tctcctcata gtttattcat tttatttatt tattttttgt ttattcgcta
     8641 ttttgtattt tagtttttaa ttagaaattt tattagaatc gttttgaact agtttttatg
     8701 cactctaaca ttaatattaa atcataacta ttatttttat tataatttat ttgtgtttat
     8761 ctttttgtgt agctttcatg tgtttttatt ttagttttat tatgtttgca ttgaattttt
     8821 tggactagtt tatataacga tatacagaaa agccatgaca acaaatgttt ttataatacc
     8881 tattaagtat aagctaatca gatttccact attctgttca aagttaacat tattagccat
     8941 tctgatggat catttgacat ttgtcatatg tctttatgaa cactgctctt tcagatgcca
     9001 cttttaaagc tgttttttac agtaaatgaa cgtttattat tatgacatta tttcctaagg
     9061 cctctagagt acagcaggat gtggcgtgtg cattaccgtg taattatgct gactgtaaat
     9121 gtgcatttat agactgtcag catgtctgac tgtgcgctgt tgtgattggt cctgcacaga
     9181 ccaag
//
