LOCUS       sequence-224779-        9546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab18a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9546)
  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 9546)
  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-rab18a
FEATURES             Location/Qualifiers
     source          1..9546
                     /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(1370..1390)
                     /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(1398..1414)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1430..1564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1454..1473)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1508..1527
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(2003..2190,2191..2202)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2236..2252)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2236..2252)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2289..2309)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2289..2307)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2328..2344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2341..2363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2352..2368
                     /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(2376..2382,2383..2400,2401..2406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2421..2442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2559..2576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2730..3318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2810..2829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3489..4280,4281..4349))
                     /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     4112..4131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4350..4454)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4617..4638)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4987..4998,4999..5161)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5534..5550
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5570..5590
                     /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 ggtgaatgat gatatattga ccactctgaa
      781 aatactgatc attggcgaaa gtggagtagg caaatccagt cttcttttga gatttactga
      841 tgatacattt gatgcagaaa ttggtgcaac aataggtgtg gatttcaagg tgaaaacgct
      901 ggctgtagat ggaaaccgag caaaactagc aatctgggat acagcaggtc aagaacgttt
      961 tcggacttta acacccagtt actacagagg agctcagggt gtcattctgg tgtatgatgt
     1021 ctctaggcga gagacatttg ctaagctgga taactggttg agtgagctgg agacgtactg
     1081 tacaagaaat gatcttgtca agatgctagt gggaaacaaa atcgataagg atgatcgtga
     1141 gattgagcga gaggagggcc tgaagttcgc aagaaaacac tccatgcttt tcatagagtc
     1201 cagtgctaaa acaagggatg gtgtccagtg tgcctttgag gagctggtgg agaaaatcct
     1261 gcagactcca ggcttgtggg agagtatgca taagtcaaga ggagttgccc tgtctgaact
     1321 ctctgaaagt agtcaaggag gctgcggggc gtactgctca cttctctaac aactttatta
     1381 tacatagttg ataattcact ggccgtcgtt ttacggtacc atcgatgatg atccagacat
     1441 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1501 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1561 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1621 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc ctctagatca
     1681 gatctgcgaa gatacggcca cgggtgctct tgatcctgtg gctgattttg gactgtgctg
     1741 ctcgcagctg ctgatgaatc acatacttcc tccattttct tccactgatt gactgttata
     1801 atttccctaa tttccaggtc aaggtgctgt gcattgtggt aatagatgtg acatgacgtc
     1861 acttccaaag gaccaatgaa catgtctgac caatttcata taatgtgaaa acgattttca
     1921 taggcagaat aaataacatt taaattaaac tgggcatcag cgcaattcaa ttggtttggt
     1981 aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa
     2041 taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta
     2101 aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag
     2161 tacagtaatc aagtaaaatt actcaagtac tttacacctc tggttcttga ccccctacct
     2221 tcagcaagcc cagcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc
     2281 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2341 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2401 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2461 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2521 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2581 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2641 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2701 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2761 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2821 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2881 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2941 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3001 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3061 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3121 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3181 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3241 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3301 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3361 aacgaaaact cacgttaagg cattttggtc atgagattat caaaaaggat cttcacctag
     3421 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3481 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3541 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3601 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3661 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3721 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3781 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3841 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3901 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3961 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4021 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4081 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4141 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4201 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4261 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4321 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4381 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4441 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     4501 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4561 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4621 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4681 tctatcaggg cgatggccca cgctggctaa gaactcatca gcctccccgg tccatctacc
     4741 cacgtaccaa tgcaccaatt ggccacaatg acggctacta catggtgcca ttccttcctc
     4801 tttataggaa tggagactac ctcctgtcca acaatgctct tggatacgag tacgcctacc
     4861 tgttggaccc aggtcattgc acaacaccag aaatgccctc tgatctgcaa aagacgtgaa
     4921 tatctgttca gacacccata tccactctgt tccacacagg tcagaggttt gtccaggagt
     4981 tcttgacaga ggtgtaaaaa gtactcaaaa attttactca agtgaaagta caagtactta
     5041 gggaaaattt tactcaatta aaagtaaaag tatctggcta gaatcttact tgagtaaaag
     5101 taaaaaagta ctccattaaa attgtacttg agtattaagg aagtaaaagt aaaagcaaga
     5161 aagaaaacta gagattcttg tttaagcttt taatctcaaa aaacattaaa tgaaatgcat
     5221 acaaggtttt atcctgcttt agaactgttt gtatttaatt atcaaactat aagacagaca
     5281 atctaatgcc agtacacgct actcaaagtt gtaaaacctc agatttaact tcagtagaag
     5341 ctgattctca aaattgttag tgtcaagcct agctcttttg gggctgaaaa gcaatcctgc
     5401 agtgctgaaa agcctctcac aggcagccga tgcgggaaga ggtgtattag tcttgataga
     5461 gaggctgcaa atagcaggaa acgtgagcag agactccctg gtgtctgaaa cacaggccag
     5521 atgggccctc gagcaggaaa cagctatgac catgattacg ccaagctatc aactttgtat
     5581 agaaaagttg gcacgcgcag attccgtgtg ggcctggtaa tatcttacca ttttaagcta
     5641 ttttaagatt agggaaaaaa attaaataaa actactttaa aattactttc cttaaaaagc
     5701 aacatattta tttagttttt gtttaactgg ctcaagattg taatatttta cttttacatg
     5761 tttttttaag tacctaaatg tgttcaaaag taaagtatgt tattgttttt tttttttaat
     5821 gttagctaag ctttaaattg taatatatta cttttaaaag tgacttttta aaaatgtaaa
     5881 taaaaaagta actataaatg tgtaatgcat gttatttctg tgggttttta attttcatct
     5941 caactgggct tacaatttta acataataaa aatcaaatca aatcaaatca ggcccttatt
     6001 tgtcacatac actcttgtat actgaaataa actaaatgtt gagaaaaatt taaatacacc
     6061 aaaataacgc ataccttttt tgtaattacc taagtaaaac atttagttcc ttttttaagt
     6121 gactcaagat tgtaatctat taattgtaaa agataaaact agtgatcaat ctgacgtgct
     6181 tctatgcagc ctgtgagatt aattgtgatg tttaattatt aaacaattta aatggatcaa
     6241 acatttcaca tgtgcttatt gtgtccaaca aaaccagcag aagcacttaa aaatattgca
     6301 tgacaacatg agtttttaat gtaaattata tatacttcct aaacgaagtt aatcatattt
     6361 ctaaaataat aatgtcattg aaatatttaa ataaatgata agctttagtt gattcaacat
     6421 aaaattgaaa gcaataagga cttttttttt ttacagcggt gaagcaaaag catgtaataa
     6481 tcatataata taataaataa ataaataaat aatggctctt ggtgcaaata tgaacacttt
     6541 aaataaatat agaataaata tgatcaataa gtcataacga tgtgttttat gttgattata
     6601 atttaaacat gtttctaacg caagttttaa aaactgtttt aaagtattat tttaagttat
     6661 aagatcaatt tgactcaaca ttttgtagtg aaaatatgtt gtgtaaaatt aatataatat
     6721 acactataaa aatgcagggt tccacacaat acattaatgt tgtcccaaca caaatcgatt
     6781 aagttaacaa atatatgtgg attgaacata aaaaattaag ttgtcccaat gaaatctcaa
     6841 gaattgtgtt gtttcagctc attttaaata agtagtttta acaagtaaaa aataacatat
     6901 ttttgagtgt gtatatatat atattacaaa atataataat ataaaatgta aaagatataa
     6961 accatgctac atatatttaa gagctatttt gcacattact tttttgcaag ccattttagt
     7021 gtttttttta caaatatata ttagctaaat atacaatatc ttaaatatac aattaatcag
     7081 acatgcaatt aatgcattta acctacataa aaatacatga tcaacatatg ttttatgctg
     7141 attacaacta aatcaaacgt aatgaaatta atttataaaa gcgatttaaa tatgtcaata
     7201 ttttaagtta aaatcacttg aaacctacta taaatgaact taaattaaaa aaatctcctt
     7261 taattccaat gcaacagtgt tgttttttca gctgaggacc cccggtgcag atggttggcc
     7321 cccggcccgc tattctaagg aagcctttct taattttagg gcggcgctgc atgcgctcag
     7381 atcagattca cgggtggaga gggttatttt ctctttttta ttttttcttg cctgctcata
     7441 tttgggcctg tgtgaacccc aggctccgcc tcggccccca ggagccgctc ttatttaaag
     7501 gggatcctcg ccttcagtct tcgtcccagt ggaagcatca gtcttgtgat aacaacttgg
     7561 tgagtcctac agaccagcgc tcgactctag gctttaaatg cattgatttc tctggattag
     7621 ctaccgggag agttttaact ttgcttatta gtcttgtgct cgaactgcta ggtttaaaga
     7681 gtgttttctc taatgcatgc atgaagcgac agctgaaaac cagaaataga gcgtttgtcc
     7741 tcctgttaaa ctctagcctt tctttggcat ttgtcagtcc gagtcttcag ctgccggcag
     7801 gagattgggt ttcagtaaca cttgctgagc acaaactaca cgtgctttct tcagatttaa
     7861 ttcattatta atcattctag atttaaatga tgttcttact cattgctgat tattttagcc
     7921 ttgcggagca gattaaatga gcacgtgttt cattaagatt gatttaaata taattaatat
     7981 ttcaccactg tagttttaaa attgccttct aagtcatggt tgatttttaa taggtatcgg
     8041 ctctaattag attactattt tggtttttaa atgttatttc tatttcattt gtaggctcgt
     8101 ttttaatgtg attttattag attctagttt taaaatcatc tcttagcctt aatgagcaga
     8161 ctgaatgaac atgtttcatt aagattaact tcattataat gaattattta atcattctag
     8221 tgggttataa atcagctcta tttaaattaa ttgtcgtgtt tatttctatt taatttttga
     8281 ttattttttc agggtcgttt ttatgtgatt ttttcccgtt tctaatttta aactaattgt
     8341 ttgattttta tatgattcga ctctttttaa atgaattgtt ttgctttttt tattgagcat
     8401 atatatatta tacaatttca atcaacaatt gttgtgcttt ttaaatgttg tttctatttc
     8461 atttctaggg tcgattttat tcgattctag ttttaaaatc gtgattttta acttctaacc
     8521 ttactgagca gactgaatga acatgtttca tcaagattaa cttcattata gttaattatt
     8581 gaatcgtgct agtggtttat aaatcagctc tatttaaatt aatcatcatc attatttcta
     8641 tttaattttt attatttctt taggattatt tttatgtgat ttttttcacg ttctagtttt
     8701 aaactcaatg ctcattactg ctcacccttg ctgagcagtc tttcgttaag attaaattaa
     8761 ttaataacca ttctagtttt aaaacagtgt cttaaggcat agctgatatt ttacaagttt
     8821 actatttaat atatctcctt tcttttttta gggtcgattt catctgattt gattaatttt
     8881 tattatcttc actaagcttt ttgttctagg ttttattctc agtattatta ctgttatatt
     8941 tgaactgaat ttctcctcat agtttattca ttttatttat ttattttttg tttattcgct
     9001 attttgtatt ttagttttta attagaaatt ttattagaat cgttttgaac tagtttttat
     9061 gcactctaac attaatatta aatcataact attattttta ttataattta tttgtgttta
     9121 tctttttgtg tagctttcat gtgtttttat tttagtttta ttatgtttgc attgaatttt
     9181 ttggactagt ttatataacg atatacagaa aagccatgac aacaaatgtt tttataatac
     9241 ctattaagta taagctaatc agatttccac tattctgttc aaagttaaca ttattagcca
     9301 ttctgatgga tcatttgaca tttgtcatat gtctttatga acactgctct ttcagatgcc
     9361 acttttaaag ctgtttttta cagtaaatga acgtttatta ttatgacatt atttcctaag
     9421 gcctctagag tacagcagga tgtggcgtgt gcattaccgt gtaattatgc tgactgtaaa
     9481 tgtgcattta tagactgtca gcatgtctga ctgtgcgctg ttgtgattgg tcctgcacag
     9541 accaag
//
