LOCUS       sequence-206469-        9522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to dominant 
            negative CDC42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9522)
  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 9522)
  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-CDC42-DN
FEATURES             Location/Qualifiers
     source          1..9522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(514..525,526..688)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    5092..5116
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      5117..5126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5123..5818
                     /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(5820..5844)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6419..6439)
                     /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(6447..6463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6479..6613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6503..6522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6557..6576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(7052..7239,7240..7251)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7285..7301)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7285..7301)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7338..7358)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7338..7356)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7377..7393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7390..7412)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7401..7417
                     /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(7425..7431,7432..7449,7450..7455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7470..7491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7608..7625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7779..8367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7859..7878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8538..9329,9330..9398))
                     /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     9161..9180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9399..9503)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttggca cgcgcagatt ccgtgtgggc
     1141 ctggtaatat cttaccattt taagctattt taagattagg gaaaaaaatt aaataaaact
     1201 actttaaaat tactttcctt aaaaagcaac atatttattt agtttttgtt taactggctc
     1261 aagattgtaa tattttactt ttacatgttt tttttaagta cctaaatgtg ttcaaaagta
     1321 aagtatgtta ttgttttttt ttttttaatg ttagctaagc tttaaattgt aatatattac
     1381 ttttaaaagt gactttttaa aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg
     1441 ttatttctgt gggtttttta attttcatct caactgggct tacaatttta acataataaa
     1501 aatcaaatca aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa
     1561 actaaatgtt gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc
     1621 taagtaaaac atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa
     1681 agataaaact agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg
     1741 tttaattatt aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca
     1801 aaaccagcag aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata
     1861 tatacttcct aaacgaagtt aatcatattt tctaaaataa taatgtcatt gaaatattta
     1921 aataaatgat aanctttagt tgattcaaca taaaattgaa agcaataagg actttttttt
     1981 ttttacagcg gtgaagcaaa agcntgnaaa taatcatata aatataataa ataaataaat
     2041 aaaataatgg ttcttggtgc aaatatgaac actttaaata aatatagaat aaatatgatc
     2101 aataagtcat aacgacgtgt tttatgttga ttataattta aacatgtttc taacgcaagt
     2161 tttaaaaact gttttaaagt attattttaa gttataagat caatttgact caacattttg
     2221 tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc aaggttccac
     2281 acaattcatt catgttgtcc caacacaaat cgattaagtt aacaaatata tgtggattga
     2341 acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc agctcatttt
     2401 aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata tatatatatt
     2461 tatatattac aaaatataat aatataaaat gtaaaagata taaaccatgc tacatatatt
     2521 taagagctat tttgcacatt acttttttgc aagccatttg agtgtttttt tacaaatata
     2581 tattagctaa atatacaata tcttaaatat acaattaatc agacatgcaa aaaatgcatt
     2641 taacctacat aaaaatacat gatcaacata tgttttatgc tgattacaac taaatcaaac
     2701 gtaatgaaat taatttataa aagcgattta aatatgtcaa tattttaagt taaaatcact
     2761 tgaaacctac tataaatgaa cttaaattta aaaaatctcc tttaattcca atgcagcact
     2821 gttgtttttt cagctgagga cccccggtgc agatggttgg cccccggccc gctattctaa
     2881 ggaagccttt cttaatttta gggcggcgct gcatgcgctc agatcagatt cacgggtgga
     2941 gagggttatt ttctcttttt tattttttct tgcctgctca tatttgggcc tgtgtgaacc
     3001 ccaggctccg cctcggcccc caggagccgc tcttatttaa aggggatcct cgccttcagt
     3061 cttcgtccca gtggaagcat cagtcttgtg ataacaactt ggtgagtcct acagaccagc
     3121 gctcgactct aggctttaaa tgcattgatt tctctggatt agctaccggg agagttttaa
     3181 ctttgcttat tagtcttgtg cttgaactgc taggtttaaa gagggttttc tctaatgcat
     3241 gcatgaagcg acagctgaaa accagaaata gagcgtttgt cctcctgtta aactctagcc
     3301 tttctttggc atttgtcagt ccgagtcttc agctgccggc aggagattgg gtttcagtaa
     3361 cacttgctga gcacaaacta cacgtgcttc cttcagattt aattcattat taatcattct
     3421 agatttaaat gatcttctta ctcattgctg attattttag ccttgcggag cagattaaat
     3481 gagcacgtgt ttattaagat tgatttaaat ataattaata tttcaccact gtagttttaa
     3541 aattgccttc taagtcatgg ttgattttta ataggtatcg gctctaatta gattactatt
     3601 ttggttttta aatgttattt ctatttcatt tgtaggctcg tttttaatgt gattttatta
     3661 gattctagtt ttaaaatcat cttttagcct tactgagcag actgaatgaa catgtttcat
     3721 taagattaac ttcattatag ttaattattt aatcattcta gtggattata aatcagctct
     3781 atttaaattg attgtcgtgt ttatttctat ttgatttttg attatttttt aaggtcgttt
     3841 ttatgtgatt tttccccgat tctaatttta aactaattgt ttgattttta tatgattcga
     3901 ctctttttaa atgaattgtt ttgctttttt tattgagcat atatatatat tatacaattt
     3961 caatcaacaa ttgttgtgct ttttaaatgt tgtttctatt tcatttctag ggtcgatttt
     4021 attcgattct agttttaaaa tcgtgatttc taactcctaa ccttactgag cagactgaat
     4081 gaacatgttt catcaagatt aacttcatta tagttaatta ttgaatcggg ctagtggttt
     4141 ataaatcagc tctattataa attaattgtc atcattattt ctatttaatt tttattattt
     4201 ttttaggatt atttttaagg gatttttttc acgttctagt tttaaactca atgctcatta
     4261 ctgctcaccc ttgctgagca gtctttcgtt aagattaaat taattaataa cccattctag
     4321 ttttaaaaca gtgtcttaag gcatagctga tattttacaa gtttactatt taatatatct
     4381 ccatttcttt ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa
     4441 gctttttgtt ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc
     4501 ctcatagttt attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt
     4561 ttttaattag aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa
     4621 tattaaatca taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct
     4681 ttcatgtgtt tttattttag ttttattatg tttgcattga attttttgga ctagtttata
     4741 taacgatata cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc
     4801 taatcagatt tccactattc tgttcaaagt taacattatt agccattctg atggatcatt
     4861 tgacatttgt catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt
     4921 ttttacagta aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag
     4981 caggatgtgg cgtgtgcatt actgtgtaat tatgctgact gtaaatgtgc atttatagac
     5041 tgtcagcatg tctgactgtg cgctgttgtg atttgtcctg cacagaccaa ggcaagtttg
     5101 tacaaaaaag caggctgcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa
     5161 gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg
     5221 caagccctac gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc
     5281 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca
     5341 cacccagggc atccccgact tctttaagca gtccttcccc gagggcttca catgggagag
     5401 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     5461 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt
     5521 gatgcagaag aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg
     5581 cggcctggaa ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg
     5641 caacttgaag accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt
     5701 ctactatgtg gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga
     5761 gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagaga
     5821 cccagctttc ttgtacaaag tggctcagac gatcaagtgc gtcgttgttg gtgatggtgc
     5881 agtgggtaaa aactgtctat taatctccta tacaactaat aaattcccct ccgaatatgt
     5941 accaacggtc tttgataact atgcagttac ggttatgatt ggtggtgagc cgtacaccct
     6001 ggggctgttt gatactgcag gtcaggaaga ttacgataga ttacgacccc tgagttaccc
     6061 tcagacagat gtcttcttag tctgtttctc agttgtttcg ccttcttcat ttgaaaacgt
     6121 caaagaaaag tgggtacccg agatcaccca ccactgtcca aagacccctt tcctgctggt
     6181 ggggacgcag attgatctga gagatgatcc ttctactatc gaaaagcttg ccaagaacaa
     6241 acagaaaccc atcactccag agacggcaga gaagctggcc cgcgatctga aggctgtcaa
     6301 atacgtggaa tgctccgttc tgacgcagaa aggcctaaag aacgtatttg atgaggcgat
     6361 tttggcagcc ctggagcccc cagagcccaa gaagaagcgt aaatgtgtgc tgctatgaca
     6421 actttattat acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga
     6481 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     6541 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     6601 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     6661 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc
     6721 tctagatcag atctgcgaag atacggccac gggtgctctt gatcctgtgg ctgattttgg
     6781 actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt ccactgattg
     6841 actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta atagatgtga
     6901 catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat aatgtgaaaa
     6961 cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc gcaattcaat
     7021 tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt
     7081 gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa
     7141 gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata
     7201 atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac
     7261 cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc accgcggtgg
     7321 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     7381 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7441 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7501 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7561 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7621 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7681 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7741 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7801 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7861 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7921 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7981 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8041 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8101 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8161 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8221 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8281 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8341 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8401 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8461 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8521 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8581 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8641 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8701 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8761 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8821 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8881 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8941 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9001 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9061 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9121 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9181 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9241 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9301 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9361 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9421 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9481 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tg
//
