LOCUS       sequence-219918-        9209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the CaaX 
            domain from Hras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9209)
  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 9209)
  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-CaaX(tH)
FEATURES             Location/Qualifiers
     source          1..9209
                     /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"
     polyA_signal    847..981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1033..1053)
                     /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(1061..1077)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1093..1227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(1666..1853,1854..1865)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1899..1915)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1899..1915)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1952..1972)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1952..1970)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1991..2007)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2004..2026)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2015..2031
                     /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(2039..2045,2046..2063,2064..2069))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2084..2105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2222..2239)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2393..2981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2473..2492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3152..3943,3944..4012))
                     /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     3775..3794
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4013..4117)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4280..4301)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4650..4661,4662..4824)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5197..5213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5233..5253
                     /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 gggaaagctg aaccctcctg atgagagtgg
      781 ccccggctgc atgagctgca agtgtgtgct ctcctgatct agaactatag tgagtcgtat
      841 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
      901 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
      961 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1021 ggggaggtgt ggcaacttta ttatacatag ttgataattc actggccgtc gttttacggt
     1081 accatcgatg atgatccaga catgataaga tacattgatg agtttggaca aaccacaact
     1141 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1201 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1261 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1321 gctgattatg atcctctaga tcagatctgc gaagatacgg ccacgggtgc tcttgatcct
     1381 gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact tcctccattt
     1441 tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc tgtgcattgt
     1501 ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct gaccaatttc
     1561 atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta aactgggcat
     1621 cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg atctccaaaa
     1681 aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa
     1741 aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa
     1801 aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag tactttacac
     1861 ctctggttct tgacccccta ccttcagcaa gcccagcaga tccactagtt ctagagcggc
     1921 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     1981 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2041 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2101 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2161 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2221 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2281 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2341 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2401 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2461 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2521 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2581 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2641 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2701 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2761 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2821 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2881 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2941 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3001 ctacggggtc tgacgctcag tggaacgaaa actcacgtta aggcattttg gtcatgagat
     3061 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3121 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3181 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3241 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3301 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3361 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3421 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3481 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3541 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3601 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3661 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3721 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3781 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3841 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3901 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3961 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4021 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4081 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4141 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4201 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4261 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4321 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccacgctggc taagaactca
     4381 tcagcctccc cggtccatct acccacgtac caatgcacca attggccaca atgacggcta
     4441 ctacatggtg ccattccttc ctctttatag gaatggagac tacctcctgt ccaacaatgc
     4501 tcttggatac gagtacgcct acctgttgga cccaggtcat tgcacaacac cagaaatgcc
     4561 ctctgatctg caaaagacgt gaatatctgt tcagacaccc atatccactc tgttccacac
     4621 aggtcagagg tttgtccagg agttcttgac agaggtgtaa aaagtactca aaaattttac
     4681 tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg
     4741 ctagaatctt acttgagtaa aagtaaaaaa gtactccatt aaaattgtac ttgagtatta
     4801 aggaagtaaa agtaaaagca agaaagaaaa ctagagattc ttgtttaagc ttttaatctc
     4861 aaaaaacatt aaatgaaatg catacaaggt tttatcctgc tttagaactg tttgtattta
     4921 attatcaaac tataagacag acaatctaat gccagtacac gctactcaaa gttgtaaaac
     4981 ctcagattta acttcagtag aagctgattc tcaaaattgt tagtgtcaag cctagctctt
     5041 ttggggctga aaagcaatcc tgcagtgctg aaaagcctct cacaggcagc cgatgcggga
     5101 agaggtgtat tagtcttgat agagaggctg caaatagcag gaaacgtgag cagagactcc
     5161 ctggtgtctg aaacacaggc cagatgggcc ctcgagcagg aaacagctat gaccatgatt
     5221 acgccaagct atcaactttg tatagaaaag ttggcacgcg cagattccgt gtgggcctgg
     5281 taatatctta ccattttaag ctattttaag attagggaaa aaaattaaat aaaactactt
     5341 taaaattact ttccttaaaa agcaacatat ttatttagtt tttgtttaac tggctcaaga
     5401 ttgtaatatt ttacttttac atgttttttt aagtacctaa atgtgttcaa aagtaaagta
     5461 tgttattgtt tttttttttt aatgttagct aagctttaaa ttgtaatata ttacttttaa
     5521 aagtgacttt ttaaaaatgt aaataaaaaa gtaactataa atgtgtaatg catgttattt
     5581 ctgtgggttt ttaattttca tctcaactgg gcttacaatt ttaacataat aaaaatcaaa
     5641 tcaaatcaaa tcaggccctt atttgtcaca tacactcttg tatactgaaa taaactaaat
     5701 gttgagaaaa atttaaatac accaaaataa cgcatacctt ttttgtaatt acctaagtaa
     5761 aacatttagt tcctttttta agtgactcaa gattgtaatc tattaattgt aaaagataaa
     5821 actagtgatc aatctgacgt gcttctatgc agcctgtgag attaattgtg atgtttaatt
     5881 attaaacaat ttaaatggat caaacatttc acatgtgctt attgtgtcca acaaaaccag
     5941 cagaagcact taaaaatatt gcatgacaac atgagttttt aatgtaaatt atatatactt
     6001 cctaaacgaa gttaatcata tttctaaaat aataatgtca ttgaaatatt taaataaatg
     6061 ataagcttta gttgattcaa cataaaattg aaagcaataa ggactttttt tttttacagc
     6121 ggtgaagcaa aagcatgtaa taatcatata atataataaa taaataaata aataatggct
     6181 cttggtgcaa atatgaacac tttaaataaa tatagaataa atatgatcaa taagtcataa
     6241 cgatgtgttt tatgttgatt ataatttaaa catgtttcta acgcaagttt taaaaactgt
     6301 tttaaagtat tattttaagt tataagatca atttgactca acattttgta gtgaaaatat
     6361 gttgtgtaaa attaatataa tatacactat aaaaatgcag ggttccacac aatacattaa
     6421 tgttgtccca acacaaatcg attaagttaa caaatatatg tggattgaac ataaaaaatt
     6481 aagttgtccc aatgaaatct caagaattgt gttgtttcag ctcattttaa ataagtagtt
     6541 ttaacaagta aaaaataaca tatttttgag tgtgtatata tatatattac aaaatataat
     6601 aatataaaat gtaaaagata taaaccatgc tacatatatt taagagctat tttgcacatt
     6661 acttttttgc aagccatttt agtgtttttt ttacaaatat atattagcta aatatacaat
     6721 atcttaaata tacaattaat cagacatgca attaatgcat ttaacctaca taaaaataca
     6781 tgatcaacat atgttttatg ctgattacaa ctaaatcaaa cgtaatgaaa ttaatttata
     6841 aaagcgattt aaatatgtca atattttaag ttaaaatcac ttgaaaccta ctataaatga
     6901 acttaaatta aaaaaatctc ctttaattcc aatgcaacag tgttgttttt tcagctgagg
     6961 acccccggtg cagatggttg gcccccggcc cgctattcta aggaagcctt tcttaatttt
     7021 agggcggcgc tgcatgcgct cagatcagat tcacgggtgg agagggttat tttctctttt
     7081 ttattttttc ttgcctgctc atatttgggc ctgtgtgaac cccaggctcc gcctcggccc
     7141 ccaggagccg ctcttattta aaggggatcc tcgccttcag tcttcgtccc agtggaagca
     7201 tcagtcttgt gataacaact tggtgagtcc tacagaccag cgctcgactc taggctttaa
     7261 atgcattgat ttctctggat tagctaccgg gagagtttta actttgctta ttagtcttgt
     7321 gctcgaactg ctaggtttaa agagtgtttt ctctaatgca tgcatgaagc gacagctgaa
     7381 aaccagaaat agagcgtttg tcctcctgtt aaactctagc ctttctttgg catttgtcag
     7441 tccgagtctt cagctgccgg caggagattg ggtttcagta acacttgctg agcacaaact
     7501 acacgtgctt tcttcagatt taattcatta ttaatcattc tagatttaaa tgatgttctt
     7561 actcattgct gattatttta gccttgcgga gcagattaaa tgagcacgtg tttcattaag
     7621 attgatttaa atataattaa tatttcacca ctgtagtttt aaaattgcct tctaagtcat
     7681 ggttgatttt taataggtat cggctctaat tagattacta ttttggtttt taaatgttat
     7741 ttctatttca tttgtaggct cgtttttaat gtgattttat tagattctag ttttaaaatc
     7801 atctcttagc cttaatgagc agactgaatg aacatgtttc attaagatta acttcattat
     7861 aatgaattat ttaatcattc tagtgggtta taaatcagct ctatttaaat taattgtcgt
     7921 gtttatttct atttaatttt tgattatttt ttcagggtcg tttttatgtg attttttccc
     7981 gtttctaatt ttaaactaat tgtttgattt ttatatgatt cgactctttt taaatgaatt
     8041 gttttgcttt ttttattgag catatatata ttatacaatt tcaatcaaca attgttgtgc
     8101 tttttaaatg ttgtttctat ttcatttcta gggtcgattt tattcgattc tagttttaaa
     8161 atcgtgattt ttaacttcta accttactga gcagactgaa tgaacatgtt tcatcaagat
     8221 taacttcatt atagttaatt attgaatcgt gctagtggtt tataaatcag ctctatttaa
     8281 attaatcatc atcattattt ctatttaatt tttattattt ctttaggatt atttttatgt
     8341 gatttttttc acgttctagt tttaaactca atgctcatta ctgctcaccc ttgctgagca
     8401 gtctttcgtt aagattaaat taattaataa ccattctagt tttaaaacag tgtcttaagg
     8461 catagctgat attttacaag tttactattt aatatatctc ctttcttttt ttagggtcga
     8521 tttcatctga tttgattaat ttttattatc ttcactaagc tttttgttct aggttttatt
     8581 ctcagtatta ttactgttat atttgaactg aatttctcct catagtttat tcattttatt
     8641 tatttatttt ttgtttattc gctattttgt attttagttt ttaattagaa attttattag
     8701 aatcgttttg aactagtttt tatgcactct aacattaata ttaaatcata actattattt
     8761 ttattataat ttatttgtgt ttatcttttt gtgtagcttt catgtgtttt tattttagtt
     8821 ttattatgtt tgcattgaat tttttggact agtttatata acgatataca gaaaagccat
     8881 gacaacaaat gtttttataa tacctattaa gtataagcta atcagatttc cactattctg
     8941 ttcaaagtta acattattag ccattctgat ggatcatttg acatttgtca tatgtcttta
     9001 tgaacactgc tctttcagat gccactttta aagctgtttt ttacagtaaa tgaacgttta
     9061 ttattatgac attatttcct aaggcctcta gagtacagca ggatgtggcg tgtgcattac
     9121 cgtgtaatta tgctgactgt aaatgtgcat ttatagactg tcagcatgtc tgactgtgcg
     9181 ctgttgtgat tggtcctgca cagaccaag
//
