LOCUS       sequence-224786-        9556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab18b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9556)
  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 9556)
  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-rab18b
FEATURES             Location/Qualifiers
     source          1..9556
                     /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(1380..1400)
                     /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(1408..1424)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1440..1574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1464..1483)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1518..1537
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(2013..2200,2201..2212)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2246..2262)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2246..2262)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2338..2354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2351..2373)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2362..2378
                     /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(2386..2392,2393..2410,2411..2416))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2431..2452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2569..2586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2740..3328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2820..2839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3499..4290,4291..4359))
                     /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     4122..4141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4360..4464)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4627..4648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4997..5008,5009..5171)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5544..5560
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5580..5600
                     /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 ggtggacgac gacgtgttga cgaccctgaa
      781 gattttaata atcggagaga gcggagtcgg taaatcgagt ttgctcctgc gtttcacaga
      841 tgacaccttc gacccagaat tagcagcaac catcggtgtg gacttcaaag tcaaaactat
      901 tgcgatcgat ggcaaccgag caaaactggc tatatgggat accgcaggtc aggagaggtt
      961 cagaactcta acaccaagct actacagagg ggcccagggt gtcattttgg tgtatgatgt
     1021 tacaaagcgg gacacgttca caaagcttga gaactggctc aatgagctgg agacatactg
     1081 tacacgcaac gacctggtta aaatgctggt ggggaacaaa atcgacaagg acaaccgtga
     1141 agtggaccgg aacgaaggct tgaagtttgc acggaaacac tctatgctct ttatcgaggc
     1201 cagcgcgaag acgcgggacg gagttcagtg tgcctttgag gagctggtgg agaagatcct
     1261 gcagacgccg gggctctggg aaagcagcat ccagaaccat ggtgtccagc tgtctgataa
     1321 tgaaccccag cgtcaaggag cctgcggcgg atactgctca ctggtttaag gatccaccgc
     1381 aactttatta tacatagttg ataattcact ggccgtcgtt ttacggtacc atcgatgatg
     1441 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1501 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1561 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1621 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     1681 ctctagatca gatctgcgaa gatacggcca cgggtgctct tgatcctgtg gctgattttg
     1741 gactgtgctg ctcgcagctg ctgatgaatc acatacttcc tccattttct tccactgatt
     1801 gactgttata atttccctaa tttccaggtc aaggtgctgt gcattgtggt aatagatgtg
     1861 acatgacgtc acttccaaag gaccaatgaa catgtctgac caatttcata taatgtgaaa
     1921 acgattttca taggcagaat aaataacatt taaattaaac tgggcatcag cgcaattcaa
     1981 ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt
     2041 tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta
     2101 agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat
     2161 aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc tggttcttga
     2221 ccccctacct tcagcaagcc cagcagatcc actagttcta gagcggccgc caccgcggtg
     2281 gagctccagc ttttgttccc tttagtgagg gttrmttgcr mgcttggcgt aatcatggtc
     2341 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2401 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2461 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2521 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3181 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3601 gggcttacca tctggcccca gtgctgcaat gataccgcga gayccacgct caccggctcc
     3661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4381 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4441 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     4501 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4561 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4621 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4681 cgaaaaaccg tctatcaggg cgatggccca cgctggctaa gaactcatca gcctccccgg
     4741 tccatctacc cacgtaccaa tgcaccaatt ggccacaatg acggctacta catggtgcca
     4801 ttccttcctc tttataggaa tggagactac ctcctgtcca acaatgctct tggatacgag
     4861 tacgcctacc tgttggaccc aggtcattgc acaacaccag aaatgccctc tgatctgcaa
     4921 aagacgtgaa tatctgttca gacacccata tccactctgt tccacacagg tcagaggttt
     4981 gtccaggagt tcttgacaga ggtgtaaaaa gtactcaaaa attttactca agtgaaagta
     5041 caagtactta gggaaaattt tactcaatta aaagtaaaag tatctggcta gaatcttact
     5101 tgagtaaaag taaaaaagta ctccattaaa attgtacttg agtattaagg aagtaaaagt
     5161 aaaagcaaga aagaaaacta gagattcttg tttaagcttt taatctcaaa aaacattaaa
     5221 tgaaatgcat acaaggtttt atcctgcttt agaactgttt gtatttaatt atcaaactat
     5281 aagacagaca atctaatgcc agtacacgct actcaaagtt gtaaaacctc agatttaact
     5341 tcagtagaag ctgattctca aaattgttag tgtcaagcct agctcttttg gggctgaaaa
     5401 gcaatcctgc agtgctgaaa agcctctcac aggcagccga tgcgggaaga ggtgtattag
     5461 tcttgataga gaggctgcaa atagcaggaa acgtgagcag agactccctg gtgtctgaaa
     5521 cacaggccag atgggccctc gagcaggaaa cagctatgac catgattacg ccaagctatc
     5581 aactttgtat agaaaagttg gcacgcgcag attccgtgtg ggcctggtaa tatcttacca
     5641 ttttaagcta ttttaagatt agggaaaaaa attaaataaa actactttaa aattactttc
     5701 cttaaaaagc aacatattta tttagttttt gtttaactgg ctcaagattg taatatttta
     5761 cttttacatg tttttttaag tacctaaatg tgttcaaaag taaagtatgt tattgttttt
     5821 tttttttaat gttagctaag ctttaaattg taatatatta cttttaaaag tgacttttta
     5881 aaaatgtaaa taaaaaagta actataaatg tgtaatgcat gttatttctg tgggttttta
     5941 attttcatct caactgggct tacaatttta acataataaa aatcaaatca aatcaaatca
     6001 ggcccttatt tgtcacatac actcttgtat actgaaataa actaaatgtt gagaaaaatt
     6061 taaatacacc aaaataacgc ataccttttt tgtaattacc taagtaaaac atttagttcc
     6121 ttttttaagt gactcaagat tgtaatctat taattgtaaa agataaaact agtgatcaat
     6181 ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg tttaattatt aaacaattta
     6241 aatggatcaa acatttcaca tgtgcttatt gtgtccaaca aaaccagcag aagcacttaa
     6301 aaatattgca tgacaacatg agtttttaat gtaaattata tatacttcct aaacgaagtt
     6361 aatcatattt ctaaaataat aatgtcattg aaatatttaa ataaatgata agctttagtt
     6421 gattcaacat aaaattgaaa gcaataagga cttttttttt ttacagcggt gaagcaaaag
     6481 catgtaataa tcatataata taataaataa ataaataaat aatggctctt ggtgcaaata
     6541 tgaacacttt aaataaatat agaataaata tgatcaataa gtcataacga tgtgttttat
     6601 gttgattata atttaaacat gtttctaacg caagttttaa aaactgtttt aaagtattat
     6661 tttaagttat aagatcaatt tgactcaaca ttttgtagtg aaaatatgtt gtgtaaaatt
     6721 aatataatat acactataaa aatgcagggt tccacacaat acattaatgt tgtcccaaca
     6781 caaatcgatt aagttaacaa atatatgtgg attgaacata aaaaattaag ttgtcccaat
     6841 gaaatctcaa gaattgtgtt gtttcagctc attttaaata agtagtttta acaagtaaaa
     6901 aataacatat ttttgagtgt gtatatatat atattacaaa atataataat ataaaatgta
     6961 aaagatataa accatgctac atatatttaa gagctatttt gcacattact tttttgcaag
     7021 ccattttagt gtttttttta caaatatata ttagctaaat atacaatatc ttaaatatac
     7081 aattaatcag acatgcaatt aatgcattta acctacataa aaatacatga tcaacatatg
     7141 ttttatgctg attacaacta aatcaaacgt aatgaaatta atttataaaa gcgatttaaa
     7201 tatgtcaata ttttaagtta aaatcacttg aaacctacta taaatgaact taaattaaaa
     7261 aaatctcctt taattccaat gcaacagtgt tgttttttca gctgaggacc cccggtgcag
     7321 atggttggcc cccggcccgc tattctaagg aagcctttct taattttagg gcggcgctgc
     7381 atgcgctcag atcagattca cgggtggaga gggttatttt ctctttttta ttttttcttg
     7441 cctgctcata tttgggcctg tgtgaacccc aggctccgcc tcggccccca ggagccgctc
     7501 ttatttaaag gggatcctcg ccttcagtct tcgtcccagt ggaagcatca gtcttgtgat
     7561 aacaacttgg tgagtcctac agaccagcgc tcgactctag gctttaaatg cattgatttc
     7621 tctggattag ctaccgggag agttttaact ttgcttatta gtcttgtgct cgaactgcta
     7681 ggtttaaaga gtgttttctc taatgcatgc atgaagcgac agctgaaaac cagaaataga
     7741 gcgtttgtcc tcctgttaaa ctctagcctt tctttggcat ttgtcagtcc gagtcttcag
     7801 ctgccggcag gagattgggt ttcagtaaca cttgctgagc acaaactaca cgtgctttct
     7861 tcagatttaa ttcattatta atcattctag atttaaatga tgttcttact cattgctgat
     7921 tattttagcc ttgcggagca gattaaatga gcacgtgttt cattaagatt gatttaaata
     7981 taattaatat ttcaccactg tagttttaaa attgccttct aagtcatggt tgatttttaa
     8041 taggtatcgg ctctaattag attactattt tggtttttaa atgttatttc tatttcattt
     8101 gtaggctcgt ttttaatgtg attttattag attctagttt taaaatcatc tcttagcctt
     8161 aatgagcaga ctgaatgaac atgtttcatt aagattaact tcattataat gaattattta
     8221 atcattctag tgggttataa atcagctcta tttaaattaa ttgtcgtgtt tatttctatt
     8281 taatttttga ttattttttc agggtcgttt ttatgtgatt ttttcccgtt tctaatttta
     8341 aactaattgt ttgattttta tatgattcga ctctttttaa atgaattgtt ttgctttttt
     8401 tattgagcat atatatatta tacaatttca atcaacaatt gttgtgcttt ttaaatgttg
     8461 tttctatttc atttctaggg tcgattttat tcgattctag ttttaaaatc gtgattttta
     8521 acttctaacc ttactgagca gactgaatga acatgtttca tcaagattaa cttcattata
     8581 gttaattatt gaatcgtgct agtggtttat aaatcagctc tatttaaatt aatcatcatc
     8641 attatttcta tttaattttt attatttctt taggattatt tttatgtgat ttttttcacg
     8701 ttctagtttt aaactcaatg ctcattactg ctcacccttg ctgagcagtc tttcgttaag
     8761 attaaattaa ttaataacca ttctagtttt aaaacagtgt cttaaggcat agctgatatt
     8821 ttacaagttt actatttaat atatctcctt tcttttttta gggtcgattt catctgattt
     8881 gattaatttt tattatcttc actaagcttt ttgttctagg ttttattctc agtattatta
     8941 ctgttatatt tgaactgaat ttctcctcat agtttattca ttttatttat ttattttttg
     9001 tttattcgct attttgtatt ttagttttta attagaaatt ttattagaat cgttttgaac
     9061 tagtttttat gcactctaac attaatatta aatcataact attattttta ttataattta
     9121 tttgtgttta tctttttgtg tagctttcat gtgtttttat tttagtttta ttatgtttgc
     9181 attgaatttt ttggactagt ttatataacg atatacagaa aagccatgac aacaaatgtt
     9241 tttataatac ctattaagta taagctaatc agatttccac tattctgttc aaagttaaca
     9301 ttattagcca ttctgatgga tcatttgaca tttgtcatat gtctttatga acactgctct
     9361 ttcagatgcc acttttaaag ctgtttttta cagtaaatga acgtttatta ttatgacatt
     9421 atttcctaag gcctctagag tacagcagga tgtggcgtgt gcattaccgt gtaattatgc
     9481 tgactgtaaa tgtgcattta tagactgtca gcatgtctga ctgtgcgctg ttgtgattgg
     9541 tcctgcacag accaag
//
