LOCUS       sequence-221287-        9555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab6bb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9555)
  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 9555)
  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-rab6bb
FEATURES             Location/Qualifiers
     source          1..9555
                     /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(1379..1399)
                     /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(1407..1423)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1439..1573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1463..1482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1517..1536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(2012..2199,2200..2211)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2245..2261)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2245..2261)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2298..2318)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2298..2316)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2337..2353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2350..2372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2361..2377
                     /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(2385..2391,2392..2409,2410..2415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2430..2451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2568..2585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2739..3327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2819..2838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3498..4289,4290..4358))
                     /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     4121..4140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4359..4463)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4626..4647)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4996..5007,5008..5170)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5543..5559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5579..5599
                     /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 ggtgtccatg ggaagtgact ttgggaaccc
      781 tttacggaaa ttcaaacttg tttttttagg ggagcaaagc gttgggaaga catcgctgat
      841 aacaaggttc atgtatgaca gttttgacaa cacttatcag gcaactattg gcattgactt
      901 tttatcaaaa accatgtatc tggaggaccg aactgtgagg ttacagctat gggacactgc
      961 agggcaggag cgttttagga gtcttatccc gagctacatt agagactcca cagtggctgt
     1021 ggtggtgtat gatatcacaa atatcaattc attccagcta acctctaaat ggattgatga
     1081 tgtcagaaca gagaggggaa gtgatgtcat tatcatgctg gtgggcaata aaacagatct
     1141 agaggaaaag aggcaaatca ctatagagga gggggagcag cgggccaaag agttgaatgt
     1201 gatgtttatc gagactagtg caaagaccgg cagtaatgtt aaacagttgt ttcggcgagt
     1261 ggccgctgct ttgcctggta tggagagctt ggatgataat aacaaagaag gaatgattga
     1321 cattaagctg gacaaacagc cggatcctcc ggccactgaa agcggctgct cctgttagca
     1381 actttattat acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga
     1441 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1501 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1561 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1621 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc
     1681 tctagatcag atctgcgaag atacggccac gggtgctctt gatcctgtgg ctgattttgg
     1741 actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt ccactgattg
     1801 actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta atagatgtga
     1861 catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat aatgtgaaaa
     1921 cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc gcaattcaat
     1981 tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt
     2041 gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa
     2101 gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata
     2161 atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac
     2221 cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc accgcggtgg
     2281 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     2341 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2401 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2461 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3361 gctcagtgga acgaaaactc acgttaaggc attttggtca tgagattatc aaaaaggatc
     3421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     4501 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4561 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4621 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4681 gaaaaaccgt ctatcagggc gatggcccac gctggctaag aactcatcag cctccccggt
     4741 ccatctaccc acgtaccaat gcaccaattg gccacaatga cggctactac atggtgccat
     4801 tccttcctct ttataggaat ggagactacc tcctgtccaa caatgctctt ggatacgagt
     4861 acgcctacct gttggaccca ggtcattgca caacaccaga aatgccctct gatctgcaaa
     4921 agacgtgaat atctgttcag acacccatat ccactctgtt ccacacaggt cagaggtttg
     4981 tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac
     5041 aagtacttag ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt
     5101 gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga gtattaagga agtaaaagta
     5161 aaagcaagaa agaaaactag agattcttgt ttaagctttt aatctcaaaa aacattaaat
     5221 gaaatgcata caaggtttta tcctgcttta gaactgtttg tatttaatta tcaaactata
     5281 agacagacaa tctaatgcca gtacacgcta ctcaaagttg taaaacctca gatttaactt
     5341 cagtagaagc tgattctcaa aattgttagt gtcaagccta gctcttttgg ggctgaaaag
     5401 caatcctgca gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag gtgtattagt
     5461 cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg tgtctgaaac
     5521 acaggccaga tgggccctcg agcaggaaac agctatgacc atgattacgc caagctatca
     5581 actttgtata gaaaagttgg cacgcgcaga ttccgtgtgg gcctggtaat atcttaccat
     5641 tttaagctat tttaagatta gggaaaaaaa ttaaataaaa ctactttaaa attactttcc
     5701 ttaaaaagca acatatttat ttagtttttg tttaactggc tcaagattgt aatattttac
     5761 ttttacatgt ttttttaagt acctaaatgt gttcaaaagt aaagtatgtt attgtttttt
     5821 ttttttaatg ttagctaagc tttaaattgt aatatattac ttttaaaagt gactttttaa
     5881 aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg ttatttctgt gggtttttaa
     5941 ttttcatctc aactgggctt acaattttaa cataataaaa atcaaatcaa atcaaatcag
     6001 gcccttattt gtcacataca ctcttgtata ctgaaataaa ctaaatgttg agaaaaattt
     6061 aaatacacca aaataacgca tacctttttt gtaattacct aagtaaaaca tttagttcct
     6121 tttttaagtg actcaagatt gtaatctatt aattgtaaaa gataaaacta gtgatcaatc
     6181 tgacgtgctt ctatgcagcc tgtgagatta attgtgatgt ttaattatta aacaatttaa
     6241 atggatcaaa catttcacat gtgcttattg tgtccaacaa aaccagcaga agcacttaaa
     6301 aatattgcat gacaacatga gtttttaatg taaattatat atacttccta aacgaagtta
     6361 atcatatttc taaaataata atgtcattga aatatttaaa taaatgataa gctttagttg
     6421 attcaacata aaattgaaag caataaggac tttttttttt tacagcggtg aagcaaaagc
     6481 atgtaataat catataatat aataaataaa taaataaata atggctcttg gtgcaaatat
     6541 gaacacttta aataaatata gaataaatat gatcaataag tcataacgat gtgttttatg
     6601 ttgattataa tttaaacatg tttctaacgc aagttttaaa aactgtttta aagtattatt
     6661 ttaagttata agatcaattt gactcaacat tttgtagtga aaatatgttg tgtaaaatta
     6721 atataatata cactataaaa atgcagggtt ccacacaata cattaatgtt gtcccaacac
     6781 aaatcgatta agttaacaaa tatatgtgga ttgaacataa aaaattaagt tgtcccaatg
     6841 aaatctcaag aattgtgttg tttcagctca ttttaaataa gtagttttaa caagtaaaaa
     6901 ataacatatt tttgagtgtg tatatatata tattacaaaa tataataata taaaatgtaa
     6961 aagatataaa ccatgctaca tatatttaag agctattttg cacattactt ttttgcaagc
     7021 cattttagtg ttttttttac aaatatatat tagctaaata tacaatatct taaatataca
     7081 attaatcaga catgcaatta atgcatttaa cctacataaa aatacatgat caacatatgt
     7141 tttatgctga ttacaactaa atcaaacgta atgaaattaa tttataaaag cgatttaaat
     7201 atgtcaatat tttaagttaa aatcacttga aacctactat aaatgaactt aaattaaaaa
     7261 aatctccttt aattccaatg caacagtgtt gttttttcag ctgaggaccc ccggtgcaga
     7321 tggttggccc ccggcccgct attctaagga agcctttctt aattttaggg cggcgctgca
     7381 tgcgctcaga tcagattcac gggtggagag ggttattttc tcttttttat tttttcttgc
     7441 ctgctcatat ttgggcctgt gtgaacccca ggctccgcct cggcccccag gagccgctct
     7501 tatttaaagg ggatcctcgc cttcagtctt cgtcccagtg gaagcatcag tcttgtgata
     7561 acaacttggt gagtcctaca gaccagcgct cgactctagg ctttaaatgc attgatttct
     7621 ctggattagc taccgggaga gttttaactt tgcttattag tcttgtgctc gaactgctag
     7681 gtttaaagag tgttttctct aatgcatgca tgaagcgaca gctgaaaacc agaaatagag
     7741 cgtttgtcct cctgttaaac tctagccttt ctttggcatt tgtcagtccg agtcttcagc
     7801 tgccggcagg agattgggtt tcagtaacac ttgctgagca caaactacac gtgctttctt
     7861 cagatttaat tcattattaa tcattctaga tttaaatgat gttcttactc attgctgatt
     7921 attttagcct tgcggagcag attaaatgag cacgtgtttc attaagattg atttaaatat
     7981 aattaatatt tcaccactgt agttttaaaa ttgccttcta agtcatggtt gatttttaat
     8041 aggtatcggc tctaattaga ttactatttt ggtttttaaa tgttatttct atttcatttg
     8101 taggctcgtt tttaatgtga ttttattaga ttctagtttt aaaatcatct cttagcctta
     8161 atgagcagac tgaatgaaca tgtttcatta agattaactt cattataatg aattatttaa
     8221 tcattctagt gggttataaa tcagctctat ttaaattaat tgtcgtgttt atttctattt
     8281 aatttttgat tattttttca gggtcgtttt tatgtgattt tttcccgttt ctaattttaa
     8341 actaattgtt tgatttttat atgattcgac tctttttaaa tgaattgttt tgcttttttt
     8401 attgagcata tatatattat acaatttcaa tcaacaattg ttgtgctttt taaatgttgt
     8461 ttctatttca tttctagggt cgattttatt cgattctagt tttaaaatcg tgatttttaa
     8521 cttctaacct tactgagcag actgaatgaa catgtttcat caagattaac ttcattatag
     8581 ttaattattg aatcgtgcta gtggtttata aatcagctct atttaaatta atcatcatca
     8641 ttatttctat ttaattttta ttatttcttt aggattattt ttatgtgatt tttttcacgt
     8701 tctagtttta aactcaatgc tcattactgc tcacccttgc tgagcagtct ttcgttaaga
     8761 ttaaattaat taataaccat tctagtttta aaacagtgtc ttaaggcata gctgatattt
     8821 tacaagttta ctatttaata tatctccttt ctttttttag ggtcgatttc atctgatttg
     8881 attaattttt attatcttca ctaagctttt tgttctaggt tttattctca gtattattac
     8941 tgttatattt gaactgaatt tctcctcata gtttattcat tttatttatt tattttttgt
     9001 ttattcgcta ttttgtattt tagtttttaa ttagaaattt tattagaatc gttttgaact
     9061 agtttttatg cactctaaca ttaatattaa atcataacta ttatttttat tataatttat
     9121 ttgtgtttat ctttttgtgt agctttcatg tgtttttatt ttagttttat tatgtttgca
     9181 ttgaattttt tggactagtt tatataacga tatacagaaa agccatgaca acaaatgttt
     9241 ttataatacc tattaagtat aagctaatca gatttccact attctgttca aagttaacat
     9301 tattagccat tctgatggat catttgacat ttgtcatatg tctttatgaa cactgctctt
     9361 tcagatgcca cttttaaagc tgttttttac agtaaatgaa cgtttattat tatgacatta
     9421 tttcctaagg cctctagagt acagcaggat gtggcgtgtg cattaccgtg taattatgct
     9481 gactgtaaat gtgcatttat agactgtcag catgtctgac tgtgcgctgt tgtgattggt
     9541 cctgcacaga ccaag
//
