LOCUS       sequence-224839-        9174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the CaaX 
            domain from Kras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9174)
  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 9174)
  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(tK)
FEATURES             Location/Qualifiers
     source          1..9174
                     /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    812..946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(998..1018)
                     /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(1026..1042)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1058..1192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(1631..1818,1819..1830)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1864..1880)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1864..1880)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1917..1937)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1917..1935)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1956..1972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1969..1991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1980..1996
                     /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(2004..2010,2011..2028,2029..2034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2049..2070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2187..2204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2358..2946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2438..2457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3117..3908,3909..3977))
                     /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     3740..3759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3978..4082)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4245..4266)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4615..4626,4627..4789)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5162..5178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5198..5218
                     /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 gggaaagaag aagaagaaga agagcaagac
      781 caagtgcgtg atcatgtgat cgtattacgt agatccagac atgataagat acattgatga
      841 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      901 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      961 cattcatttt atgtttcagg ttcaggggga ggtgtggcaa ctttattata catagttgat
     1021 aattcactgg ccgtcgtttt acggtaccat cgatgatgat ccagacatga taagatacat
     1081 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1141 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1201 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1261 gtaaaacctc tacaaatgtg gtatggctga ttatgatcct ctagatcaga tctgcgaaga
     1321 tacggccacg ggtgctcttg atcctgtggc tgattttgga ctgtgctgct cgcagctgct
     1381 gatgaatcac atacttcctc cattttcttc cactgattga ctgttataat ttccctaatt
     1441 tccaggtcaa ggtgctgtgc attgtggtaa tagatgtgac atgacgtcac ttccaaagga
     1501 ccaatgaaca tgtctgacca atttcatata atgtgaaaac gattttcata ggcagaataa
     1561 ataacattta aattaaactg ggcatcagcg caattcaatt ggtttggtaa tagcaaggga
     1621 aaatagaatg aagtgatctc caaaaaataa gtactttttg actgtaaata aaattgtaag
     1681 gagtaaaaag tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt
     1741 tttaattgta ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa
     1801 gtaaaattac tcaagtactt tacacctctg gttcttgacc ccctaccttc agcaagccca
     1861 gcagatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     1921 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1981 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2041 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2101 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2161 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2221 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2281 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2341 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2401 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2461 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2521 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2581 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2701 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2761 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2821 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2881 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2941 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3001 cgttaaggca ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4081 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4141 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4201 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4261 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4321 atggcccacg ctggctaaga actcatcagc ctccccggtc catctaccca cgtaccaatg
     4381 caccaattgg ccacaatgac ggctactaca tggtgccatt ccttcctctt tataggaatg
     4441 gagactacct cctgtccaac aatgctcttg gatacgagta cgcctacctg ttggacccag
     4501 gtcattgcac aacaccagaa atgccctctg atctgcaaaa gacgtgaata tctgttcaga
     4561 cacccatatc cactctgttc cacacaggtc agaggtttgt ccaggagttc ttgacagagg
     4621 tgtaaaaagt actcaaaaat tttactcaag tgaaagtaca agtacttagg gaaaatttta
     4681 ctcaattaaa agtaaaagta tctggctaga atcttacttg agtaaaagta aaaaagtact
     4741 ccattaaaat tgtacttgag tattaaggaa gtaaaagtaa aagcaagaaa gaaaactaga
     4801 gattcttgtt taagctttta atctcaaaaa acattaaatg aaatgcatac aaggttttat
     4861 cctgctttag aactgtttgt atttaattat caaactataa gacagacaat ctaatgccag
     4921 tacacgctac tcaaagttgt aaaacctcag atttaacttc agtagaagct gattctcaaa
     4981 attgttagtg tcaagcctag ctcttttggg gctgaaaagc aatcctgcag tgctgaaaag
     5041 cctctcacag gcagccgatg cgggaagagg tgtattagtc ttgatagaga ggctgcaaat
     5101 agcaggaaac gtgagcagag actccctggt gtctgaaaca caggccagat gggccctcga
     5161 gcaggaaaca gctatgacca tgattacgcc aagctatcaa ctttgtatag aaaagttggc
     5221 acgcgcagat tccgtgtggg cctggtaata tcttaccatt ttaagctatt ttaagattag
     5281 ggaaaaaaat taaataaaac tactttaaaa ttactttcct taaaaagcaa catatttatt
     5341 tagtttttgt ttaactggct caagattgta atattttact tttacatgtt tttttaagta
     5401 cctaaatgtg ttcaaaagta aagtatgtta ttgttttttt tttttaatgt tagctaagct
     5461 ttaaattgta atatattact tttaaaagtg actttttaaa aatgtaaata aaaaagtaac
     5521 tataaatgtg taatgcatgt tatttctgtg ggtttttaat tttcatctca actgggctta
     5581 caattttaac ataataaaaa tcaaatcaaa tcaaatcagg cccttatttg tcacatacac
     5641 tcttgtatac tgaaataaac taaatgttga gaaaaattta aatacaccaa aataacgcat
     5701 accttttttg taattaccta agtaaaacat ttagttcctt ttttaagtga ctcaagattg
     5761 taatctatta attgtaaaag ataaaactag tgatcaatct gacgtgcttc tatgcagcct
     5821 gtgagattaa ttgtgatgtt taattattaa acaatttaaa tggatcaaac atttcacatg
     5881 tgcttattgt gtccaacaaa accagcagaa gcacttaaaa atattgcatg acaacatgag
     5941 tttttaatgt aaattatata tacttcctaa acgaagttaa tcatatttct aaaataataa
     6001 tgtcattgaa atatttaaat aaatgataag ctttagttga ttcaacataa aattgaaagc
     6061 aataaggact tttttttttt acagcggtga agcaaaagca tgtaataatc atataatata
     6121 ataaataaat aaataaataa tggctcttgg tgcaaatatg aacactttaa ataaatatag
     6181 aataaatatg atcaataagt cataacgatg tgttttatgt tgattataat ttaaacatgt
     6241 ttctaacgca agttttaaaa actgttttaa agtattattt taagttataa gatcaatttg
     6301 actcaacatt ttgtagtgaa aatatgttgt gtaaaattaa tataatatac actataaaaa
     6361 tgcagggttc cacacaatac attaatgttg tcccaacaca aatcgattaa gttaacaaat
     6421 atatgtggat tgaacataaa aaattaagtt gtcccaatga aatctcaaga attgtgttgt
     6481 ttcagctcat tttaaataag tagttttaac aagtaaaaaa taacatattt ttgagtgtgt
     6541 atatatatat attacaaaat ataataatat aaaatgtaaa agatataaac catgctacat
     6601 atatttaaga gctattttgc acattacttt tttgcaagcc attttagtgt tttttttaca
     6661 aatatatatt agctaaatat acaatatctt aaatatacaa ttaatcagac atgcaattaa
     6721 tgcatttaac ctacataaaa atacatgatc aacatatgtt ttatgctgat tacaactaaa
     6781 tcaaacgtaa tgaaattaat ttataaaagc gatttaaata tgtcaatatt ttaagttaaa
     6841 atcacttgaa acctactata aatgaactta aattaaaaaa atctccttta attccaatgc
     6901 aacagtgttg ttttttcagc tgaggacccc cggtgcagat ggttggcccc cggcccgcta
     6961 ttctaaggaa gcctttctta attttagggc ggcgctgcat gcgctcagat cagattcacg
     7021 ggtggagagg gttattttct cttttttatt ttttcttgcc tgctcatatt tgggcctgtg
     7081 tgaaccccag gctccgcctc ggcccccagg agccgctctt atttaaaggg gatcctcgcc
     7141 ttcagtcttc gtcccagtgg aagcatcagt cttgtgataa caacttggtg agtcctacag
     7201 accagcgctc gactctaggc tttaaatgca ttgatttctc tggattagct accgggagag
     7261 ttttaacttt gcttattagt cttgtgctcg aactgctagg tttaaagagt gttttctcta
     7321 atgcatgcat gaagcgacag ctgaaaacca gaaatagagc gtttgtcctc ctgttaaact
     7381 ctagcctttc tttggcattt gtcagtccga gtcttcagct gccggcagga gattgggttt
     7441 cagtaacact tgctgagcac aaactacacg tgctttcttc agatttaatt cattattaat
     7501 cattctagat ttaaatgatg ttcttactca ttgctgatta ttttagcctt gcggagcaga
     7561 ttaaatgagc acgtgtttca ttaagattga tttaaatata attaatattt caccactgta
     7621 gttttaaaat tgccttctaa gtcatggttg atttttaata ggtatcggct ctaattagat
     7681 tactattttg gtttttaaat gttatttcta tttcatttgt aggctcgttt ttaatgtgat
     7741 tttattagat tctagtttta aaatcatctc ttagccttaa tgagcagact gaatgaacat
     7801 gtttcattaa gattaacttc attataatga attatttaat cattctagtg ggttataaat
     7861 cagctctatt taaattaatt gtcgtgttta tttctattta atttttgatt attttttcag
     7921 ggtcgttttt atgtgatttt ttcccgtttc taattttaaa ctaattgttt gatttttata
     7981 tgattcgact ctttttaaat gaattgtttt gcttttttta ttgagcatat atatattata
     8041 caatttcaat caacaattgt tgtgcttttt aaatgttgtt tctatttcat ttctagggtc
     8101 gattttattc gattctagtt ttaaaatcgt gatttttaac ttctaacctt actgagcaga
     8161 ctgaatgaac atgtttcatc aagattaact tcattatagt taattattga atcgtgctag
     8221 tggtttataa atcagctcta tttaaattaa tcatcatcat tatttctatt taatttttat
     8281 tatttcttta ggattatttt tatgtgattt ttttcacgtt ctagttttaa actcaatgct
     8341 cattactgct cacccttgct gagcagtctt tcgttaagat taaattaatt aataaccatt
     8401 ctagttttaa aacagtgtct taaggcatag ctgatatttt acaagtttac tatttaatat
     8461 atctcctttc tttttttagg gtcgatttca tctgatttga ttaattttta ttatcttcac
     8521 taagcttttt gttctaggtt ttattctcag tattattact gttatatttg aactgaattt
     8581 ctcctcatag tttattcatt ttatttattt attttttgtt tattcgctat tttgtatttt
     8641 agtttttaat tagaaatttt attagaatcg ttttgaacta gtttttatgc actctaacat
     8701 taatattaaa tcataactat tatttttatt ataatttatt tgtgtttatc tttttgtgta
     8761 gctttcatgt gtttttattt tagttttatt atgtttgcat tgaatttttt ggactagttt
     8821 atataacgat atacagaaaa gccatgacaa caaatgtttt tataatacct attaagtata
     8881 agctaatcag atttccacta ttctgttcaa agttaacatt attagccatt ctgatggatc
     8941 atttgacatt tgtcatatgt ctttatgaac actgctcttt cagatgccac ttttaaagct
     9001 gttttttaca gtaaatgaac gtttattatt atgacattat ttcctaaggc ctctagagta
     9061 cagcaggatg tggcgtgtgc attaccgtgt aattatgctg actgtaaatg tgcatttata
     9121 gactgtcagc atgtctgact gtgcgctgtt gtgattggtc ctgcacagac caag
//
