LOCUS       sequence-224751-        9513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to Rab31.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9513)
  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 9513)
  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-Rab31
FEATURES             Location/Qualifiers
     source          1..9513
                     /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(1337..1357)
                     /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(1365..1381)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1397..1531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1421..1440)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1475..1494
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1970..2157,2158..2169)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2203..2219)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2203..2219)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2256..2276)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2256..2274)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2295..2311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2308..2330)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2319..2335
                     /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(2343..2349,2350..2367,2368..2373))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2388..2409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2526..2543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2697..3285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2777..2796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3456..4247,4248..4316))
                     /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     4079..4098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4317..4421)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4584..4605)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4954..4965,4966..5128)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5501..5517
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5537..5557
                     /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 ggtggccata agggagctaa aagtttgtct
      781 gctcggggac actggtgtgg ggaaatcgag tatcgtgtgt cggtttgttc aagatcactt
      841 tgaccacaac attagtccga caataggtgc atcattttta actaagaccg tcccctctgg
      901 aaatgaactt cacaaatttc tgatctggga cactgcagga caagagcggt ttcattccct
      961 ggctcccatg tattacagag gctcggcagc ggctgtcatt gtttatgaca tcacaaagct
     1021 tgactcgttc cagacgttaa agaagtgggt gaaagagctg aaggagcacg ggccggagga
     1081 tatagtggtg gccatcgctg gaaataagaa cgatctgggt gacatcaggg aagttccaac
     1141 taaggaggcg aaagaatttg cagagtcgat tgcagccata tttatggaga ccagtgccag
     1201 aaatgctgta aacatagagg agctgttcca aaaaatcagt cgacagatcc ctccactgga
     1261 gaacactgat gcagacagcc acgactcctt caagctgacc cgacagcctc ctccagccac
     1321 caaacgctgc tgttagcaac tttattatac atagttgata attcactggc cgtcgtttta
     1381 cggtaccatc gatgatgatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     1441 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1501 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1561 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1621 tatggctgat tatgatcctc tagatcagat ctgcgaagat acggccacgg gtgctcttga
     1681 tcctgtggct gattttggac tgtgctgctc gcagctgctg atgaatcaca tacttcctcc
     1741 attttcttcc actgattgac tgttataatt tccctaattt ccaggtcaag gtgctgtgca
     1801 ttgtggtaat agatgtgaca tgacgtcact tccaaaggac caatgaacat gtctgaccaa
     1861 tttcatataa tgtgaaaacg attttcatag gcagaataaa taacatttaa attaaactgg
     1921 gcatcagcgc aattcaattg gtttggtaat agcaagggaa aatagaatga agtgatctcc
     1981 aaaaaataag tactttttga ctgtaaataa aattgtaagg agtaaaaagt actttttttt
     2041 ctaaaaaaat gtaattaagt aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag
     2101 taaaaatccc caaaaataat acttaagtac agtaatcaag taaaattact caagtacttt
     2161 acacctctgg ttcttgaccc cctaccttca gcaagcccag cagatccact agttctagag
     2221 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2281 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2341 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2401 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2461 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2521 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2581 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2641 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3121 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3301 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaaggcat tttggtcatg
     3361 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3421 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3481 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3541 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3601 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3661 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3721 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3781 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3841 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3901 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3961 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4021 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4081 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4141 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4201 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4261 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4321 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4381 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4441 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4501 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4561 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4621 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
     4681 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
     4741 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
     4801 atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
     4861 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
     4921 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
     4981 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
     5041 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
     5101 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
     5161 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
     5221 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
     5281 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
     5341 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
     5401 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     5461 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     5521 gattacgcca agctatcaac tttgtataga aaagttggca cgcgcagatt ccgtgtgggc
     5581 ctggtaatat cttaccattt taagctattt taagattagg gaaaaaaatt aaataaaact
     5641 actttaaaat tactttcctt aaaaagcaac atatttattt agtttttgtt taactggctc
     5701 aagattgtaa tattttactt ttacatgttt ttttaagtac ctaaatgtgt tcaaaagtaa
     5761 agtatgttat tgtttttttt ttttaatgtt agctaagctt taaattgtaa tatattactt
     5821 ttaaaagtga ctttttaaaa atgtaaataa aaaagtaact ataaatgtgt aatgcatgtt
     5881 atttctgtgg gtttttaatt ttcatctcaa ctgggcttac aattttaaca taataaaaat
     5941 caaatcaaat caaatcaggc ccttatttgt cacatacact cttgtatact gaaataaact
     6001 aaatgttgag aaaaatttaa atacaccaaa ataacgcata ccttttttgt aattacctaa
     6061 gtaaaacatt tagttccttt tttaagtgac tcaagattgt aatctattaa ttgtaaaaga
     6121 taaaactagt gatcaatctg acgtgcttct atgcagcctg tgagattaat tgtgatgttt
     6181 aattattaaa caatttaaat ggatcaaaca tttcacatgt gcttattgtg tccaacaaaa
     6241 ccagcagaag cacttaaaaa tattgcatga caacatgagt ttttaatgta aattatatat
     6301 acttcctaaa cgaagttaat catatttcta aaataataat gtcattgaaa tatttaaata
     6361 aatgataagc tttagttgat tcaacataaa attgaaagca ataaggactt ttttttttta
     6421 cagcggtgaa gcaaaagcat gtaataatca tataatataa taaataaata aataaataat
     6481 ggctcttggt gcaaatatga acactttaaa taaatataga ataaatatga tcaataagtc
     6541 ataacgatgt gttttatgtt gattataatt taaacatgtt tctaacgcaa gttttaaaaa
     6601 ctgttttaaa gtattatttt aagttataag atcaatttga ctcaacattt tgtagtgaaa
     6661 atatgttgtg taaaattaat ataatataca ctataaaaat gcagggttcc acacaataca
     6721 ttaatgttgt cccaacacaa atcgattaag ttaacaaata tatgtggatt gaacataaaa
     6781 aattaagttg tcccaatgaa atctcaagaa ttgtgttgtt tcagctcatt ttaaataagt
     6841 agttttaaca agtaaaaaat aacatatttt tgagtgtgta tatatatata ttacaaaata
     6901 taataatata aaatgtaaaa gatataaacc atgctacata tatttaagag ctattttgca
     6961 cattactttt ttgcaagcca ttttagtgtt ttttttacaa atatatatta gctaaatata
     7021 caatatctta aatatacaat taatcagaca tgcaattaat gcatttaacc tacataaaaa
     7081 tacatgatca acatatgttt tatgctgatt acaactaaat caaacgtaat gaaattaatt
     7141 tataaaagcg atttaaatat gtcaatattt taagttaaaa tcacttgaaa cctactataa
     7201 atgaacttaa attaaaaaaa tctcctttaa ttccaatgca acagtgttgt tttttcagct
     7261 gaggaccccc ggtgcagatg gttggccccc ggcccgctat tctaaggaag cctttcttaa
     7321 ttttagggcg gcgctgcatg cgctcagatc agattcacgg gtggagaggg ttattttctc
     7381 ttttttattt tttcttgcct gctcatattt gggcctgtgt gaaccccagg ctccgcctcg
     7441 gcccccagga gccgctctta tttaaagggg atcctcgcct tcagtcttcg tcccagtgga
     7501 agcatcagtc ttgtgataac aacttggtga gtcctacaga ccagcgctcg actctaggct
     7561 ttaaatgcat tgatttctct ggattagcta ccgggagagt tttaactttg cttattagtc
     7621 ttgtgctcga actgctaggt ttaaagagtg ttttctctaa tgcatgcatg aagcgacagc
     7681 tgaaaaccag aaatagagcg tttgtcctcc tgttaaactc tagcctttct ttggcatttg
     7741 tcagtccgag tcttcagctg ccggcaggag attgggtttc agtaacactt gctgagcaca
     7801 aactacacgt gctttcttca gatttaattc attattaatc attctagatt taaatgatgt
     7861 tcttactcat tgctgattat tttagccttg cggagcagat taaatgagca cgtgtttcat
     7921 taagattgat ttaaatataa ttaatatttc accactgtag ttttaaaatt gccttctaag
     7981 tcatggttga tttttaatag gtatcggctc taattagatt actattttgg tttttaaatg
     8041 ttatttctat ttcatttgta ggctcgtttt taatgtgatt ttattagatt ctagttttaa
     8101 aatcatctct tagccttaat gagcagactg aatgaacatg tttcattaag attaacttca
     8161 ttataatgaa ttatttaatc attctagtgg gttataaatc agctctattt aaattaattg
     8221 tcgtgtttat ttctatttaa tttttgatta ttttttcagg gtcgttttta tgtgattttt
     8281 tcccgtttct aattttaaac taattgtttg atttttatat gattcgactc tttttaaatg
     8341 aattgttttg ctttttttat tgagcatata tatattatac aatttcaatc aacaattgtt
     8401 gtgcttttta aatgttgttt ctatttcatt tctagggtcg attttattcg attctagttt
     8461 taaaatcgtg atttttaact tctaacctta ctgagcagac tgaatgaaca tgtttcatca
     8521 agattaactt cattatagtt aattattgaa tcgtgctagt ggtttataaa tcagctctat
     8581 ttaaattaat catcatcatt atttctattt aatttttatt atttctttag gattattttt
     8641 atgtgatttt tttcacgttc tagttttaaa ctcaatgctc attactgctc acccttgctg
     8701 agcagtcttt cgttaagatt aaattaatta ataaccattc tagttttaaa acagtgtctt
     8761 aaggcatagc tgatatttta caagtttact atttaatata tctcctttct ttttttaggg
     8821 tcgatttcat ctgatttgat taatttttat tatcttcact aagctttttg ttctaggttt
     8881 tattctcagt attattactg ttatatttga actgaatttc tcctcatagt ttattcattt
     8941 tatttattta ttttttgttt attcgctatt ttgtatttta gtttttaatt agaaatttta
     9001 ttagaatcgt tttgaactag tttttatgca ctctaacatt aatattaaat cataactatt
     9061 atttttatta taatttattt gtgtttatct ttttgtgtag ctttcatgtg tttttatttt
     9121 agttttatta tgtttgcatt gaattttttg gactagttta tataacgata tacagaaaag
     9181 ccatgacaac aaatgttttt ataataccta ttaagtataa gctaatcaga tttccactat
     9241 tctgttcaaa gttaacatta ttagccattc tgatggatca tttgacattt gtcatatgtc
     9301 tttatgaaca ctgctctttc agatgccact tttaaagctg ttttttacag taaatgaacg
     9361 tttattatta tgacattatt tcctaaggcc tctagagtac agcaggatgt ggcgtgtgca
     9421 ttaccgtgta attatgctga ctgtaaatgt gcatttatag actgtcagca tgtctgactg
     9481 tgcgctgttg tgattggtcc tgcacagacc aag
//
