LOCUS       sequence-220544-        9531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9531)
  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 9531)
  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-rab13
FEATURES             Location/Qualifiers
     source          1..9531
                     /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(1355..1375)
                     /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(1383..1399)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1415..1549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1439..1458)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1493..1512
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1988..2175,2176..2187)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2221..2237)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2221..2237)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2274..2294)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2274..2292)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2313..2329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2326..2348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2337..2353
                     /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(2361..2367,2368..2385,2386..2391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2406..2427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2544..2561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2715..3303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2795..2814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3474..4265,4266..4334))
                     /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     4097..4116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4335..4439)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4602..4623)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4972..4983,4984..5146)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5519..5535
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5555..5575
                     /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 ggtggcaaag aagtacgact tcctctttaa
      781 attattgctt atcggcgata gcggtgttgg caagacatgt ttgatcatcc gtttcgcaga
      841 ggacaatttt aattcaacgt acatctcaac catcggtatt gacttcaaag tgaagaccat
      901 tgaggtggaa ggaaagaaag tgaagctgca agtctgggac acagcaggac aggagaggtt
      961 taagaccatc accactgcat actacagagg ggccatggga atcattctcg tgtatgacat
     1021 aactgatgaa aaatcctatg agaatattca gaactggatg aagagcatca aagaaaatgc
     1081 atcagcgggt gtaagtcgaa tgttactggg taacaagtgt gacattgagg ccaagaggaa
     1141 agtctcaaaa gagactggtg agaagctcgc aaaggaacat ggtattcgat tctttgagac
     1201 tagtgccaaa tcgagcatca acgttgagga gtcttttact tctcttgctc gagacatttt
     1261 gctgaagtca aataagaagc cgggtccatc tggacgtgag gtcaaactta ccagcacaga
     1321 gaaaaagtcc tcctccaaat gtcttcttct ttagcaactt tattatacat agttgataat
     1381 tcactggccg tcgttttacg gtaccatcga tgatgatcca gacatgataa gatacattga
     1441 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1501 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1561 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1621 aaacctctac aaatgtggta tggctgatta tgatcctcta gatcagatct gcgaagatac
     1681 ggccacgggt gctcttgatc ctgtggctga ttttggactg tgctgctcgc agctgctgat
     1741 gaatcacata cttcctccat tttcttccac tgattgactg ttataatttc cctaatttcc
     1801 aggtcaaggt gctgtgcatt gtggtaatag atgtgacatg acgtcacttc caaaggacca
     1861 atgaacatgt ctgaccaatt tcatataatg tgaaaacgat tttcataggc agaataaata
     1921 acatttaaat taaactgggc atcagcgcaa ttcaattggt ttggtaatag caagggaaaa
     1981 tagaatgaag tgatctccaa aaaataagta ctttttgact gtaaataaaa ttgtaaggag
     2041 taaaaagtac ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt attgattttt
     2101 aattgtactc aagtaaagta aaaatcccca aaaataatac ttaagtacag taatcaagta
     2161 aaattactca agtactttac acctctggtt cttgaccccc taccttcagc aagcccagca
     2221 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     2281 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2341 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2401 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2461 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2521 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2581 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2641 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2701 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2761 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2821 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2881 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2941 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3001 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3061 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3121 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3181 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3241 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3301 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3361 taaggcattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3421 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3901 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4141 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4321 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4381 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4441 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     4501 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4561 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4621 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4681 gcccacgctg gctaagaact catcagcctc cccggtccat ctacccacgt accaatgcac
     4741 caattggcca caatgacggc tactacatgg tgccattcct tcctctttat aggaatggag
     4801 actacctcct gtccaacaat gctcttggat acgagtacgc ctacctgttg gacccaggtc
     4861 attgcacaac accagaaatg ccctctgatc tgcaaaagac gtgaatatct gttcagacac
     4921 ccatatccac tctgttccac acaggtcaga ggtttgtcca ggagttcttg acagaggtgt
     4981 aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa aattttactc
     5041 aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa aagtactcca
     5101 ttaaaattgt acttgagtat taaggaagta aaagtaaaag caagaaagaa aactagagat
     5161 tcttgtttaa gcttttaatc tcaaaaaaca ttaaatgaaa tgcatacaag gttttatcct
     5221 gctttagaac tgtttgtatt taattatcaa actataagac agacaatcta atgccagtac
     5281 acgctactca aagttgtaaa acctcagatt taacttcagt agaagctgat tctcaaaatt
     5341 gttagtgtca agcctagctc ttttggggct gaaaagcaat cctgcagtgc tgaaaagcct
     5401 ctcacaggca gccgatgcgg gaagaggtgt attagtcttg atagagaggc tgcaaatagc
     5461 aggaaacgtg agcagagact ccctggtgtc tgaaacacag gccagatggg ccctcgagca
     5521 ggaaacagct atgaccatga ttacgccaag ctatcaactt tgtatagaaa agttggcacg
     5581 cgcagattcc gtgtgggcct ggtaatatct taccatttta agctatttta agattaggga
     5641 aaaaaattaa ataaaactac tttaaaatta ctttccttaa aaagcaacat atttatttag
     5701 tttttgttta actggctcaa gattgtaata ttttactttt acatgttttt ttaagtacct
     5761 aaatgtgttc aaaagtaaag tatgttattg tttttttttt ttaatgttag ctaagcttta
     5821 aattgtaata tattactttt aaaagtgact ttttaaaaat gtaaataaaa aagtaactat
     5881 aaatgtgtaa tgcatgttat ttctgtgggt ttttaatttt catctcaact gggcttacaa
     5941 ttttaacata ataaaaatca aatcaaatca aatcaggccc ttatttgtca catacactct
     6001 tgtatactga aataaactaa atgttgagaa aaatttaaat acaccaaaat aacgcatacc
     6061 ttttttgtaa ttacctaagt aaaacattta gttccttttt taagtgactc aagattgtaa
     6121 tctattaatt gtaaaagata aaactagtga tcaatctgac gtgcttctat gcagcctgtg
     6181 agattaattg tgatgtttaa ttattaaaca atttaaatgg atcaaacatt tcacatgtgc
     6241 ttattgtgtc caacaaaacc agcagaagca cttaaaaata ttgcatgaca acatgagttt
     6301 ttaatgtaaa ttatatatac ttcctaaacg aagttaatca tatttctaaa ataataatgt
     6361 cattgaaata tttaaataaa tgataagctt tagttgattc aacataaaat tgaaagcaat
     6421 aaggactttt tttttttaca gcggtgaagc aaaagcatgt aataatcata taatataata
     6481 aataaataaa taaataatgg ctcttggtgc aaatatgaac actttaaata aatatagaat
     6541 aaatatgatc aataagtcat aacgatgtgt tttatgttga ttataattta aacatgtttc
     6601 taacgcaagt tttaaaaact gttttaaagt attattttaa gttataagat caatttgact
     6661 caacattttg tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc
     6721 agggttccac acaatacatt aatgttgtcc caacacaaat cgattaagtt aacaaatata
     6781 tgtggattga acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc
     6841 agctcatttt aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata
     6901 tatatatatt acaaaatata ataatataaa atgtaaaaga tataaaccat gctacatata
     6961 tttaagagct attttgcaca ttactttttt gcaagccatt ttagtgtttt ttttacaaat
     7021 atatattagc taaatataca atatcttaaa tatacaatta atcagacatg caattaatgc
     7081 atttaaccta cataaaaata catgatcaac atatgtttta tgctgattac aactaaatca
     7141 aacgtaatga aattaattta taaaagcgat ttaaatatgt caatatttta agttaaaatc
     7201 acttgaaacc tactataaat gaacttaaat taaaaaaatc tcctttaatt ccaatgcaac
     7261 agtgttgttt tttcagctga ggacccccgg tgcagatggt tggcccccgg cccgctattc
     7321 taaggaagcc tttcttaatt ttagggcggc gctgcatgcg ctcagatcag attcacgggt
     7381 ggagagggtt attttctctt ttttattttt tcttgcctgc tcatatttgg gcctgtgtga
     7441 accccaggct ccgcctcggc ccccaggagc cgctcttatt taaaggggat cctcgccttc
     7501 agtcttcgtc ccagtggaag catcagtctt gtgataacaa cttggtgagt cctacagacc
     7561 agcgctcgac tctaggcttt aaatgcattg atttctctgg attagctacc gggagagttt
     7621 taactttgct tattagtctt gtgctcgaac tgctaggttt aaagagtgtt ttctctaatg
     7681 catgcatgaa gcgacagctg aaaaccagaa atagagcgtt tgtcctcctg ttaaactcta
     7741 gcctttcttt ggcatttgtc agtccgagtc ttcagctgcc ggcaggagat tgggtttcag
     7801 taacacttgc tgagcacaaa ctacacgtgc tttcttcaga tttaattcat tattaatcat
     7861 tctagattta aatgatgttc ttactcattg ctgattattt tagccttgcg gagcagatta
     7921 aatgagcacg tgtttcatta agattgattt aaatataatt aatatttcac cactgtagtt
     7981 ttaaaattgc cttctaagtc atggttgatt tttaataggt atcggctcta attagattac
     8041 tattttggtt tttaaatgtt atttctattt catttgtagg ctcgttttta atgtgatttt
     8101 attagattct agttttaaaa tcatctctta gccttaatga gcagactgaa tgaacatgtt
     8161 tcattaagat taacttcatt ataatgaatt atttaatcat tctagtgggt tataaatcag
     8221 ctctatttaa attaattgtc gtgtttattt ctatttaatt tttgattatt ttttcagggt
     8281 cgtttttatg tgattttttc ccgtttctaa ttttaaacta attgtttgat ttttatatga
     8341 ttcgactctt tttaaatgaa ttgttttgct ttttttattg agcatatata tattatacaa
     8401 tttcaatcaa caattgttgt gctttttaaa tgttgtttct atttcatttc tagggtcgat
     8461 tttattcgat tctagtttta aaatcgtgat ttttaacttc taaccttact gagcagactg
     8521 aatgaacatg tttcatcaag attaacttca ttatagttaa ttattgaatc gtgctagtgg
     8581 tttataaatc agctctattt aaattaatca tcatcattat ttctatttaa tttttattat
     8641 ttctttagga ttatttttat gtgatttttt tcacgttcta gttttaaact caatgctcat
     8701 tactgctcac ccttgctgag cagtctttcg ttaagattaa attaattaat aaccattcta
     8761 gttttaaaac agtgtcttaa ggcatagctg atattttaca agtttactat ttaatatatc
     8821 tcctttcttt ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa
     8881 gctttttgtt ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc
     8941 ctcatagttt attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt
     9001 ttttaattag aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa
     9061 tattaaatca taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct
     9121 ttcatgtgtt tttattttag ttttattatg tttgcattga attttttgga ctagtttata
     9181 taacgatata cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc
     9241 taatcagatt tccactattc tgttcaaagt taacattatt agccattctg atggatcatt
     9301 tgacatttgt catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt
     9361 ttttacagta aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag
     9421 caggatgtgg cgtgtgcatt accgtgtaat tatgctgact gtaaatgtgc atttatagac
     9481 tgtcagcatg tctgactgtg cgctgttgtg attggtcctg cacagaccaa g
//
