LOCUS       sequence-220540-        9590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab11al.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9590)
  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 9590)
  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-rab11al
FEATURES             Location/Qualifiers
     source          1..9590
                     /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(1414..1434)
                     /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(1442..1458)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1474..1608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1498..1517)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1552..1571
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(2047..2234,2235..2246)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2280..2296)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2280..2296)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2333..2353)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2333..2351)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2372..2388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2385..2407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2396..2412
                     /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(2420..2426,2427..2444,2445..2450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2465..2486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2603..2620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2774..3362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2854..2873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3533..4324,4325..4393))
                     /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     4156..4175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4394..4498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4661..4682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5031..5042,5043..5205)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5578..5594
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5614..5634
                     /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 ggtgactggt agagaggacg agtatgacta
      781 tctctttaaa gtggttctga ttggtgactc gggtgtgggc aagagtaacc tgctctctcg
      841 cttcacccgc aatgagttca acctggagag caagagcacc atcggggtgg agtttgccac
      901 gcgcagtatc catgtggagg gcaagacggt caaagctcag atctgggata cggctggaca
      961 ggagagatac agagccatca catcagcgta ttacagaggt gccgtaggag ccctgctggt
     1021 gtatgacatc gccaagcatc tgacctatga gaacgcagag cgctggctga aagagctgca
     1081 ggatcacgct gacagcaata ttgtcatcat gttagtagga aacaaaagcg acttgcgtca
     1141 tctgagggct gttcctatgg atgaagctaa agcatttgca gagaaacatg ggctgtcttt
     1201 cttggagacg tcagcgctgg actcttctaa tgttgaactt gcctttcaga ctattctcac
     1261 agaaatctac cgtatcgtgt cccagaggca gatgtcgggg cgtggagatg acagcttctc
     1321 ccccagctcc aaagtggttc ccattacggt gcagcccaca caaaattctg gcaagcagag
     1381 tgcctgctgt cagaataact gagctgccac agccaacttt attatacata gttgataatt
     1441 cactggccgt cgttttacgg taccatcgat gatgatccag acatgataag atacattgat
     1501 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1561 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1621 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1681 aacctctaca aatgtggtat ggctgattat gatcctctag atcagatctg cgaagatacg
     1741 gccacgggtg ctcttgatcc tgtggctgat tttggactgt gctgctcgca gctgctgatg
     1801 aatcacatac ttcctccatt ttcttccact gattgactgt tataatttcc ctaatttcca
     1861 ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga cgtcacttcc aaaggaccaa
     1921 tgaacatgtc tgaccaattt catataatgt gaaaacgatt ttcataggca gaataaataa
     1981 catttaaatt aaactgggca tcagcgcaat tcaattggtt tggtaatagc aagggaaaat
     2041 agaatgaagt gatctccaaa aaataagtac tttttgactg taaataaaat tgtaaggagt
     2101 aaaaagtact tttttttcta aaaaaatgta attaagtaaa agtaaaagta ttgattttta
     2161 attgtactca agtaaagtaa aaatccccaa aaataatact taagtacagt aatcaagtaa
     2221 aattactcaa gtactttaca cctctggttc ttgaccccct accttcagca agcccagcag
     2281 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     2341 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2401 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2461 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2521 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3421 aaggcatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4501 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     4561 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4621 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4681 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4741 cccacgctgg ctaagaactc atcagcctcc ccggtccatc tacccacgta ccaatgcacc
     4801 aattggccac aatgacggct actacatggt gccattcctt cctctttata ggaatggaga
     4861 ctacctcctg tccaacaatg ctcttggata cgagtacgcc tacctgttgg acccaggtca
     4921 ttgcacaaca ccagaaatgc cctctgatct gcaaaagacg tgaatatctg ttcagacacc
     4981 catatccact ctgttccaca caggtcagag gtttgtccag gagttcttga cagaggtgta
     5041 aaaagtactc aaaaatttta ctcaagtgaa agtacaagta cttagggaaa attttactca
     5101 attaaaagta aaagtatctg gctagaatct tacttgagta aaagtaaaaa agtactccat
     5161 taaaattgta cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa actagagatt
     5221 cttgtttaag cttttaatct caaaaaacat taaatgaaat gcatacaagg ttttatcctg
     5281 ctttagaact gtttgtattt aattatcaaa ctataagaca gacaatctaa tgccagtaca
     5341 cgctactcaa agttgtaaaa cctcagattt aacttcagta gaagctgatt ctcaaaattg
     5401 ttagtgtcaa gcctagctct tttggggctg aaaagcaatc ctgcagtgct gaaaagcctc
     5461 tcacaggcag ccgatgcggg aagaggtgta ttagtcttga tagagaggct gcaaatagca
     5521 ggaaacgtga gcagagactc cctggtgtct gaaacacagg ccagatgggc cctcgagcag
     5581 gaaacagcta tgaccatgat tacgccaagc tatcaacttt gtatagaaaa gttggcacgc
     5641 gcagattccg tgtgggcctg gtaatatctt accattttaa gctattttaa gattagggaa
     5701 aaaaattaaa taaaactact ttaaaattac tttccttaaa aagcaacata tttatttagt
     5761 ttttgtttaa ctggctcaag attgtaatat tttactttta catgtttttt taagtaccta
     5821 aatgtgttca aaagtaaagt atgttattgt tttttttttt taatgttagc taagctttaa
     5881 attgtaatat attactttta aaagtgactt tttaaaaatg taaataaaaa agtaactata
     5941 aatgtgtaat gcatgttatt tctgtgggtt tttaattttc atctcaactg ggcttacaat
     6001 tttaacataa taaaaatcaa atcaaatcaa atcaggccct tatttgtcac atacactctt
     6061 gtatactgaa ataaactaaa tgttgagaaa aatttaaata caccaaaata acgcatacct
     6121 tttttgtaat tacctaagta aaacatttag ttcctttttt aagtgactca agattgtaat
     6181 ctattaattg taaaagataa aactagtgat caatctgacg tgcttctatg cagcctgtga
     6241 gattaattgt gatgtttaat tattaaacaa tttaaatgga tcaaacattt cacatgtgct
     6301 tattgtgtcc aacaaaacca gcagaagcac ttaaaaatat tgcatgacaa catgagtttt
     6361 taatgtaaat tatatatact tcctaaacga agttaatcat atttctaaaa taataatgtc
     6421 attgaaatat ttaaataaat gataagcttt agttgattca acataaaatt gaaagcaata
     6481 aggacttttt ttttttacag cggtgaagca aaagcatgta ataatcatat aatataataa
     6541 ataaataaat aaataatggc tcttggtgca aatatgaaca ctttaaataa atatagaata
     6601 aatatgatca ataagtcata acgatgtgtt ttatgttgat tataatttaa acatgtttct
     6661 aacgcaagtt ttaaaaactg ttttaaagta ttattttaag ttataagatc aatttgactc
     6721 aacattttgt agtgaaaata tgttgtgtaa aattaatata atatacacta taaaaatgca
     6781 gggttccaca caatacatta atgttgtccc aacacaaatc gattaagtta acaaatatat
     6841 gtggattgaa cataaaaaat taagttgtcc caatgaaatc tcaagaattg tgttgtttca
     6901 gctcatttta aataagtagt tttaacaagt aaaaaataac atatttttga gtgtgtatat
     6961 atatatatta caaaatataa taatataaaa tgtaaaagat ataaaccatg ctacatatat
     7021 ttaagagcta ttttgcacat tacttttttg caagccattt tagtgttttt tttacaaata
     7081 tatattagct aaatatacaa tatcttaaat atacaattaa tcagacatgc aattaatgca
     7141 tttaacctac ataaaaatac atgatcaaca tatgttttat gctgattaca actaaatcaa
     7201 acgtaatgaa attaatttat aaaagcgatt taaatatgtc aatattttaa gttaaaatca
     7261 cttgaaacct actataaatg aacttaaatt aaaaaaatct cctttaattc caatgcaaca
     7321 gtgttgtttt ttcagctgag gacccccggt gcagatggtt ggcccccggc ccgctattct
     7381 aaggaagcct ttcttaattt tagggcggcg ctgcatgcgc tcagatcaga ttcacgggtg
     7441 gagagggtta ttttctcttt tttatttttt cttgcctgct catatttggg cctgtgtgaa
     7501 ccccaggctc cgcctcggcc cccaggagcc gctcttattt aaaggggatc ctcgccttca
     7561 gtcttcgtcc cagtggaagc atcagtcttg tgataacaac ttggtgagtc ctacagacca
     7621 gcgctcgact ctaggcttta aatgcattga tttctctgga ttagctaccg ggagagtttt
     7681 aactttgctt attagtcttg tgctcgaact gctaggttta aagagtgttt tctctaatgc
     7741 atgcatgaag cgacagctga aaaccagaaa tagagcgttt gtcctcctgt taaactctag
     7801 cctttctttg gcatttgtca gtccgagtct tcagctgccg gcaggagatt gggtttcagt
     7861 aacacttgct gagcacaaac tacacgtgct ttcttcagat ttaattcatt attaatcatt
     7921 ctagatttaa atgatgttct tactcattgc tgattatttt agccttgcgg agcagattaa
     7981 atgagcacgt gtttcattaa gattgattta aatataatta atatttcacc actgtagttt
     8041 taaaattgcc ttctaagtca tggttgattt ttaataggta tcggctctaa ttagattact
     8101 attttggttt ttaaatgtta tttctatttc atttgtaggc tcgtttttaa tgtgatttta
     8161 ttagattcta gttttaaaat catctcttag ccttaatgag cagactgaat gaacatgttt
     8221 cattaagatt aacttcatta taatgaatta tttaatcatt ctagtgggtt ataaatcagc
     8281 tctatttaaa ttaattgtcg tgtttatttc tatttaattt ttgattattt tttcagggtc
     8341 gtttttatgt gattttttcc cgtttctaat tttaaactaa ttgtttgatt tttatatgat
     8401 tcgactcttt ttaaatgaat tgttttgctt tttttattga gcatatatat attatacaat
     8461 ttcaatcaac aattgttgtg ctttttaaat gttgtttcta tttcatttct agggtcgatt
     8521 ttattcgatt ctagttttaa aatcgtgatt tttaacttct aaccttactg agcagactga
     8581 atgaacatgt ttcatcaaga ttaacttcat tatagttaat tattgaatcg tgctagtggt
     8641 ttataaatca gctctattta aattaatcat catcattatt tctatttaat ttttattatt
     8701 tctttaggat tatttttatg tgattttttt cacgttctag ttttaaactc aatgctcatt
     8761 actgctcacc cttgctgagc agtctttcgt taagattaaa ttaattaata accattctag
     8821 ttttaaaaca gtgtcttaag gcatagctga tattttacaa gtttactatt taatatatct
     8881 cctttctttt tttagggtcg atttcatctg atttgattaa tttttattat cttcactaag
     8941 ctttttgttc taggttttat tctcagtatt attactgtta tatttgaact gaatttctcc
     9001 tcatagttta ttcattttat ttatttattt tttgtttatt cgctattttg tattttagtt
     9061 tttaattaga aattttatta gaatcgtttt gaactagttt ttatgcactc taacattaat
     9121 attaaatcat aactattatt tttattataa tttatttgtg tttatctttt tgtgtagctt
     9181 tcatgtgttt ttattttagt tttattatgt ttgcattgaa ttttttggac tagtttatat
     9241 aacgatatac agaaaagcca tgacaacaaa tgtttttata atacctatta agtataagct
     9301 aatcagattt ccactattct gttcaaagtt aacattatta gccattctga tggatcattt
     9361 gacatttgtc atatgtcttt atgaacactg ctctttcaga tgccactttt aaagctgttt
     9421 tttacagtaa atgaacgttt attattatga cattatttcc taaggcctct agagtacagc
     9481 aggatgtggc gtgtgcatta ccgtgtaatt atgctgactg taaatgtgca tttatagact
     9541 gtcagcatgt ctgactgtgc gctgttgtga ttggtcctgc acagaccaag
//
