LOCUS       sequence-221224-        9534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab30.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9534)
  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 9534)
  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-rab30
FEATURES             Location/Qualifiers
     source          1..9534
                     /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(1358..1378)
                     /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(1386..1402)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1418..1552
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1442..1461)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1496..1515
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1991..2178,2179..2190)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2224..2240)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2224..2240)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2277..2297)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2277..2295)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2316..2332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2329..2351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2340..2356
                     /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(2364..2370,2371..2388,2389..2394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2409..2430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2547..2564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2718..3306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2798..2817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3477..4268,4269..4337))
                     /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     4100..4119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4338..4442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4605..4626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4975..4986,4987..5149)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5522..5538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5558..5578
                     /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 ggtggaggat tatgattact tattcaaaat
      781 tgtgctgata ggaaatgcag gagtgggtaa aacctgcctg gtgcgacgtt tcactcaggg
      841 gctgtttcct cctggacagg gtgccacaat tggagttgat tttatgatta aaactgtgga
      901 aataaaagga gttaaggtga agctccagat ttgggacaca gctggtcaag aaagatttcg
      961 ctctataact cagagctact accgcagtgc caatgccctg atcctcacct acgacatcac
     1021 ctgcgaagat tcgttccggt gccttcctga gtggctgagg gagattgagc agtacgccaa
     1081 caaccaagtg gtgacaatcc ttgtagggaa taagatagac ttggctgata agcgagaagt
     1141 ccttcgtcaa cgagcagagg agtttgcaga ttctcagagc atgctttatc tggagacctc
     1201 tgccaaagaa tcagacaacg tggagaagct ctttctagat ctggcctgcg aactaatccg
     1261 tgaggccaag caaaacacgt tggacaacaa cgactccact cccatgcccg gagaagggaa
     1321 aaacatcaac tatttgggct gctgcagcat gaactagcaa ctttattata catagttgat
     1381 aattcactgg ccgtcgtttt acggtaccat cgatgatgat ccagacatga taagatacat
     1441 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1501 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1561 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1621 gtaaaacctc tacaaatgtg gtatggctga ttatgatcct ctagatcaga tctgcgaaga
     1681 tacggccacg ggtgctcttg atcctgtggc tgattttgga ctgtgctgct cgcagctgct
     1741 gatgaatcac atacttcctc cattttcttc cactgattga ctgttataat ttccctaatt
     1801 tccaggtcaa ggtgctgtgc attgtggtaa tagatgtgac atgacgtcac ttccaaagga
     1861 ccaatgaaca tgtctgacca atttcatata atgtgaaaac gattttcata ggcagaataa
     1921 ataacattta aattaaactg ggcatcagcg caattcaatt ggtttggtaa tagcaaggga
     1981 aaatagaatg aagtgatctc caaaaaataa gtactttttg actgtaaata aaattgtaag
     2041 gagtaaaaag tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt
     2101 tttaattgta ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa
     2161 gtaaaattac tcaagtactt tacacctctg gttcttgacc ccctaccttc agcaagccca
     2221 gcagatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     2281 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2341 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2401 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2461 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2521 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2581 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2641 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2701 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2761 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2821 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2881 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2941 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3001 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3061 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3121 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3181 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3241 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3301 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3361 cgttaaggca ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3421 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3481 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3541 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3601 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3661 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3721 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3781 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3841 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3901 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3961 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4021 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4081 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4141 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4201 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4261 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4321 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4381 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4441 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4501 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4561 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4621 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4681 atggcccacg ctggctaaga actcatcagc ctccccggtc catctaccca cgtaccaatg
     4741 caccaattgg ccacaatgac ggctactaca tggtgccatt ccttcctctt tataggaatg
     4801 gagactacct cctgtccaac aatgctcttg gatacgagta cgcctacctg ttggacccag
     4861 gtcattgcac aacaccagaa atgccctctg atctgcaaaa gacgtgaata tctgttcaga
     4921 cacccatatc cactctgttc cacacaggtc agaggtttgt ccaggagttc ttgacagagg
     4981 tgtaaaaagt actcaaaaat tttactcaag tgaaagtaca agtacttagg gaaaatttta
     5041 ctcaattaaa agtaaaagta tctggctaga atcttacttg agtaaaagta aaaaagtact
     5101 ccattaaaat tgtacttgag tattaaggaa gtaaaagtaa aagcaagaaa gaaaactaga
     5161 gattcttgtt taagctttta atctcaaaaa acattaaatg aaatgcatac aaggttttat
     5221 cctgctttag aactgtttgt atttaattat caaactataa gacagacaat ctaatgccag
     5281 tacacgctac tcaaagttgt aaaacctcag atttaacttc agtagaagct gattctcaaa
     5341 attgttagtg tcaagcctag ctcttttggg gctgaaaagc aatcctgcag tgctgaaaag
     5401 cctctcacag gcagccgatg cgggaagagg tgtattagtc ttgatagaga ggctgcaaat
     5461 agcaggaaac gtgagcagag actccctggt gtctgaaaca caggccagat gggccctcga
     5521 gcaggaaaca gctatgacca tgattacgcc aagctatcaa ctttgtatag aaaagttggc
     5581 acgcgcagat tccgtgtggg cctggtaata tcttaccatt ttaagctatt ttaagattag
     5641 ggaaaaaaat taaataaaac tactttaaaa ttactttcct taaaaagcaa catatttatt
     5701 tagtttttgt ttaactggct caagattgta atattttact tttacatgtt tttttaagta
     5761 cctaaatgtg ttcaaaagta aagtatgtta ttgttttttt tttttaatgt tagctaagct
     5821 ttaaattgta atatattact tttaaaagtg actttttaaa aatgtaaata aaaaagtaac
     5881 tataaatgtg taatgcatgt tatttctgtg ggtttttaat tttcatctca actgggctta
     5941 caattttaac ataataaaaa tcaaatcaaa tcaaatcagg cccttatttg tcacatacac
     6001 tcttgtatac tgaaataaac taaatgttga gaaaaattta aatacaccaa aataacgcat
     6061 accttttttg taattaccta agtaaaacat ttagttcctt ttttaagtga ctcaagattg
     6121 taatctatta attgtaaaag ataaaactag tgatcaatct gacgtgcttc tatgcagcct
     6181 gtgagattaa ttgtgatgtt taattattaa acaatttaaa tggatcaaac atttcacatg
     6241 tgcttattgt gtccaacaaa accagcagaa gcacttaaaa atattgcatg acaacatgag
     6301 tttttaatgt aaattatata tacttcctaa acgaagttaa tcatatttct aaaataataa
     6361 tgtcattgaa atatttaaat aaatgataag ctttagttga ttcaacataa aattgaaagc
     6421 aataaggact tttttttttt acagcggtga agcaaaagca tgtaataatc atataatata
     6481 ataaataaat aaataaataa tggctcttgg tgcaaatatg aacactttaa ataaatatag
     6541 aataaatatg atcaataagt cataacgatg tgttttatgt tgattataat ttaaacatgt
     6601 ttctaacgca agttttaaaa actgttttaa agtattattt taagttataa gatcaatttg
     6661 actcaacatt ttgtagtgaa aatatgttgt gtaaaattaa tataatatac actataaaaa
     6721 tgcagggttc cacacaatac attaatgttg tcccaacaca aatcgattaa gttaacaaat
     6781 atatgtggat tgaacataaa aaattaagtt gtcccaatga aatctcaaga attgtgttgt
     6841 ttcagctcat tttaaataag tagttttaac aagtaaaaaa taacatattt ttgagtgtgt
     6901 atatatatat attacaaaat ataataatat aaaatgtaaa agatataaac catgctacat
     6961 atatttaaga gctattttgc acattacttt tttgcaagcc attttagtgt tttttttaca
     7021 aatatatatt agctaaatat acaatatctt aaatatacaa ttaatcagac atgcaattaa
     7081 tgcatttaac ctacataaaa atacatgatc aacatatgtt ttatgctgat tacaactaaa
     7141 tcaaacgtaa tgaaattaat ttataaaagc gatttaaata tgtcaatatt ttaagttaaa
     7201 atcacttgaa acctactata aatgaactta aattaaaaaa atctccttta attccaatgc
     7261 aacagtgttg ttttttcagc tgaggacccc cggtgcagat ggttggcccc cggcccgcta
     7321 ttctaaggaa gcctttctta attttagggc ggcgctgcat gcgctcagat cagattcacg
     7381 ggtggagagg gttattttct cttttttatt ttttcttgcc tgctcatatt tgggcctgtg
     7441 tgaaccccag gctccgcctc ggcccccagg agccgctctt atttaaaggg gatcctcgcc
     7501 ttcagtcttc gtcccagtgg aagcatcagt cttgtgataa caacttggtg agtcctacag
     7561 accagcgctc gactctaggc tttaaatgca ttgatttctc tggattagct accgggagag
     7621 ttttaacttt gcttattagt cttgtgctcg aactgctagg tttaaagagt gttttctcta
     7681 atgcatgcat gaagcgacag ctgaaaacca gaaatagagc gtttgtcctc ctgttaaact
     7741 ctagcctttc tttggcattt gtcagtccga gtcttcagct gccggcagga gattgggttt
     7801 cagtaacact tgctgagcac aaactacacg tgctttcttc agatttaatt cattattaat
     7861 cattctagat ttaaatgatg ttcttactca ttgctgatta ttttagcctt gcggagcaga
     7921 ttaaatgagc acgtgtttca ttaagattga tttaaatata attaatattt caccactgta
     7981 gttttaaaat tgccttctaa gtcatggttg atttttaata ggtatcggct ctaattagat
     8041 tactattttg gtttttaaat gttatttcta tttcatttgt aggctcgttt ttaatgtgat
     8101 tttattagat tctagtttta aaatcatctc ttagccttaa tgagcagact gaatgaacat
     8161 gtttcattaa gattaacttc attataatga attatttaat cattctagtg ggttataaat
     8221 cagctctatt taaattaatt gtcgtgttta tttctattta atttttgatt attttttcag
     8281 ggtcgttttt atgtgatttt ttcccgtttc taattttaaa ctaattgttt gatttttata
     8341 tgattcgact ctttttaaat gaattgtttt gcttttttta ttgagcatat atatattata
     8401 caatttcaat caacaattgt tgtgcttttt aaatgttgtt tctatttcat ttctagggtc
     8461 gattttattc gattctagtt ttaaaatcgt gatttttaac ttctaacctt actgagcaga
     8521 ctgaatgaac atgtttcatc aagattaact tcattatagt taattattga atcgtgctag
     8581 tggtttataa atcagctcta tttaaattaa tcatcatcat tatttctatt taatttttat
     8641 tatttcttta ggattatttt tatgtgattt ttttcacgtt ctagttttaa actcaatgct
     8701 cattactgct cacccttgct gagcagtctt tcgttaagat taaattaatt aataaccatt
     8761 ctagttttaa aacagtgtct taaggcatag ctgatatttt acaagtttac tatttaatat
     8821 atctcctttc tttttttagg gtcgatttca tctgatttga ttaattttta ttatcttcac
     8881 taagcttttt gttctaggtt ttattctcag tattattact gttatatttg aactgaattt
     8941 ctcctcatag tttattcatt ttatttattt attttttgtt tattcgctat tttgtatttt
     9001 agtttttaat tagaaatttt attagaatcg ttttgaacta gtttttatgc actctaacat
     9061 taatattaaa tcataactat tatttttatt ataatttatt tgtgtttatc tttttgtgta
     9121 gctttcatgt gtttttattt tagttttatt atgtttgcat tgaatttttt ggactagttt
     9181 atataacgat atacagaaaa gccatgacaa caaatgtttt tataatacct attaagtata
     9241 agctaatcag atttccacta ttctgttcaa agttaacatt attagccatt ctgatggatc
     9301 atttgacatt tgtcatatgt ctttatgaac actgctcttt cagatgccac ttttaaagct
     9361 gttttttaca gtaaatgaac gtttattatt atgacattat ttcctaaggc ctctagagta
     9421 cagcaggatg tggcgtgtgc attaccgtgt aattatgctg actgtaaatg tgcatttata
     9481 gactgtcagc atgtctgact gtgcgctgtt gtgattggtc ctgcacagac caag
//
