LOCUS       sequence-224840-        9202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the 5' 
            signal sequence from calreticulin and 3' KDEL ER retention sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9202)
  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 9202)
  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-KDEL-pA
FEATURES             Location/Qualifiers
     source          1..9202
                     /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"
     CDS             89..781
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     protein_bind    complement(803..827)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    829..963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1026..1046)
                     /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(1054..1070)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1086..1220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(1659..1846,1847..1858)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1892..1908)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1892..1908)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1945..1965)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1945..1963)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1984..2000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1997..2019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2008..2024
                     /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(2032..2038,2039..2056,2057..2062))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2077..2098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2215..2232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2386..2974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2466..2485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3145..3936,3937..4005))
                     /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     3768..3787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4006..4110)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4273..4294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4643..4654,4655..4817)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5190..5206
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5226..5246
                     /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 catgctgcta tccgtgccgt tgctgctcgg
       61 cctcctcggc ctggccgtcg ccctcgaggt gagcgagctg attaaggaga acatgcacat
      121 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
      181 aggcaagccc tacgagggca cccagaccat gagaatcaag gcggtcgagg gcggccctct
      241 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa
      301 ccacacccag ggcatccccg acttctttaa gcagtccttc cccgagggct tcacatggga
      361 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
      421 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc
      481 tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagaccctgt accccgctga
      541 cggcggcctg gaaggcagag ccgacatggc cctgaagctc gtgggcgggg gccacctgat
      601 ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg
      661 cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt
      721 cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacag
      781 aggatccaag gacgagctgt aaacccagct ttcttgtaca aagtggggga tccagacatg
      841 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
      901 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
      961 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1021 ttttccaact ttattataca tagttgataa ttcactggcc gtcgttttac ggtaccatcg
     1081 atgatgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1141 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1201 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1261 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     1321 atgatcctct agatcagatc tgcgaagata cggccacggg tgctcttgat cctgtggctg
     1381 attttggact gtgctgctcg cagctgctga tgaatcacat acttcctcca ttttcttcca
     1441 ctgattgact gttataattt ccctaatttc caggtcaagg tgctgtgcat tgtggtaata
     1501 gatgtgacat gacgtcactt ccaaaggacc aatgaacatg tctgaccaat ttcatataat
     1561 gtgaaaacga ttttcatagg cagaataaat aacatttaaa ttaaactggg catcagcgca
     1621 attcaattgg tttggtaata gcaagggaaa atagaatgaa gtgatctcca aaaaataagt
     1681 actttttgac tgtaaataaa attgtaagga gtaaaaagta cttttttttc taaaaaaatg
     1741 taattaagta aaagtaaaag tattgatttt taattgtact caagtaaagt aaaaatcccc
     1801 aaaaataata cttaagtaca gtaatcaagt aaaattactc aagtacttta cacctctggt
     1861 tcttgacccc ctaccttcag caagcccagc agatccacta gttctagagc ggccgccacc
     1921 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     1981 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2041 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2101 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2161 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2221 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2281 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2521 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2821 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3001 gtctgacgct cagtggaacg aaaactcacg ttaaggcatt ttggtcatga gattatcaaa
     3061 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4141 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4201 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4261 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4321 aaagggcgaa aaaccgtcta tcagggcgat ggcccacgct ggctaagaac tcatcagcct
     4381 ccccggtcca tctacccacg taccaatgca ccaattggcc acaatgacgg ctactacatg
     4441 gtgccattcc ttcctcttta taggaatgga gactacctcc tgtccaacaa tgctcttgga
     4501 tacgagtacg cctacctgtt ggacccaggt cattgcacaa caccagaaat gccctctgat
     4561 ctgcaaaaga cgtgaatatc tgttcagaca cccatatcca ctctgttcca cacaggtcag
     4621 aggtttgtcc aggagttctt gacagaggtg taaaaagtac tcaaaaattt tactcaagtg
     4681 aaagtacaag tacttaggga aaattttact caattaaaag taaaagtatc tggctagaat
     4741 cttacttgag taaaagtaaa aaagtactcc attaaaattg tacttgagta ttaaggaagt
     4801 aaaagtaaaa gcaagaaaga aaactagaga ttcttgttta agcttttaat ctcaaaaaac
     4861 attaaatgaa atgcatacaa ggttttatcc tgctttagaa ctgtttgtat ttaattatca
     4921 aactataaga cagacaatct aatgccagta cacgctactc aaagttgtaa aacctcagat
     4981 ttaacttcag tagaagctga ttctcaaaat tgttagtgtc aagcctagct cttttggggc
     5041 tgaaaagcaa tcctgcagtg ctgaaaagcc tctcacaggc agccgatgcg ggaagaggtg
     5101 tattagtctt gatagagagg ctgcaaatag caggaaacgt gagcagagac tccctggtgt
     5161 ctgaaacaca ggccagatgg gccctcgagc aggaaacagc tatgaccatg attacgccaa
     5221 gctatcaact ttgtatagaa aagttggcac gcgcagattc cgtgtgggcc tggtaatatc
     5281 ttaccatttt aagctatttt aagattaggg aaaaaaatta aataaaacta ctttaaaatt
     5341 actttcctta aaaagcaaca tatttattta gtttttgttt aactggctca agattgtaat
     5401 attttacttt tacatgtttt tttaagtacc taaatgtgtt caaaagtaaa gtatgttatt
     5461 gttttttttt tttaatgtta gctaagcttt aaattgtaat atattacttt taaaagtgac
     5521 tttttaaaaa tgtaaataaa aaagtaacta taaatgtgta atgcatgtta tttctgtggg
     5581 tttttaattt tcatctcaac tgggcttaca attttaacat aataaaaatc aaatcaaatc
     5641 aaatcaggcc cttatttgtc acatacactc ttgtatactg aaataaacta aatgttgaga
     5701 aaaatttaaa tacaccaaaa taacgcatac cttttttgta attacctaag taaaacattt
     5761 agttcctttt ttaagtgact caagattgta atctattaat tgtaaaagat aaaactagtg
     5821 atcaatctga cgtgcttcta tgcagcctgt gagattaatt gtgatgttta attattaaac
     5881 aatttaaatg gatcaaacat ttcacatgtg cttattgtgt ccaacaaaac cagcagaagc
     5941 acttaaaaat attgcatgac aacatgagtt tttaatgtaa attatatata cttcctaaac
     6001 gaagttaatc atatttctaa aataataatg tcattgaaat atttaaataa atgataagct
     6061 ttagttgatt caacataaaa ttgaaagcaa taaggacttt ttttttttac agcggtgaag
     6121 caaaagcatg taataatcat ataatataat aaataaataa ataaataatg gctcttggtg
     6181 caaatatgaa cactttaaat aaatatagaa taaatatgat caataagtca taacgatgtg
     6241 ttttatgttg attataattt aaacatgttt ctaacgcaag ttttaaaaac tgttttaaag
     6301 tattatttta agttataaga tcaatttgac tcaacatttt gtagtgaaaa tatgttgtgt
     6361 aaaattaata taatatacac tataaaaatg cagggttcca cacaatacat taatgttgtc
     6421 ccaacacaaa tcgattaagt taacaaatat atgtggattg aacataaaaa attaagttgt
     6481 cccaatgaaa tctcaagaat tgtgttgttt cagctcattt taaataagta gttttaacaa
     6541 gtaaaaaata acatattttt gagtgtgtat atatatatat tacaaaatat aataatataa
     6601 aatgtaaaag atataaacca tgctacatat atttaagagc tattttgcac attacttttt
     6661 tgcaagccat tttagtgttt tttttacaaa tatatattag ctaaatatac aatatcttaa
     6721 atatacaatt aatcagacat gcaattaatg catttaacct acataaaaat acatgatcaa
     6781 catatgtttt atgctgatta caactaaatc aaacgtaatg aaattaattt ataaaagcga
     6841 tttaaatatg tcaatatttt aagttaaaat cacttgaaac ctactataaa tgaacttaaa
     6901 ttaaaaaaat ctcctttaat tccaatgcaa cagtgttgtt ttttcagctg aggacccccg
     6961 gtgcagatgg ttggcccccg gcccgctatt ctaaggaagc ctttcttaat tttagggcgg
     7021 cgctgcatgc gctcagatca gattcacggg tggagagggt tattttctct tttttatttt
     7081 ttcttgcctg ctcatatttg ggcctgtgtg aaccccaggc tccgcctcgg cccccaggag
     7141 ccgctcttat ttaaagggga tcctcgcctt cagtcttcgt cccagtggaa gcatcagtct
     7201 tgtgataaca acttggtgag tcctacagac cagcgctcga ctctaggctt taaatgcatt
     7261 gatttctctg gattagctac cgggagagtt ttaactttgc ttattagtct tgtgctcgaa
     7321 ctgctaggtt taaagagtgt tttctctaat gcatgcatga agcgacagct gaaaaccaga
     7381 aatagagcgt ttgtcctcct gttaaactct agcctttctt tggcatttgt cagtccgagt
     7441 cttcagctgc cggcaggaga ttgggtttca gtaacacttg ctgagcacaa actacacgtg
     7501 ctttcttcag atttaattca ttattaatca ttctagattt aaatgatgtt cttactcatt
     7561 gctgattatt ttagccttgc ggagcagatt aaatgagcac gtgtttcatt aagattgatt
     7621 taaatataat taatatttca ccactgtagt tttaaaattg ccttctaagt catggttgat
     7681 ttttaatagg tatcggctct aattagatta ctattttggt ttttaaatgt tatttctatt
     7741 tcatttgtag gctcgttttt aatgtgattt tattagattc tagttttaaa atcatctctt
     7801 agccttaatg agcagactga atgaacatgt ttcattaaga ttaacttcat tataatgaat
     7861 tatttaatca ttctagtggg ttataaatca gctctattta aattaattgt cgtgtttatt
     7921 tctatttaat ttttgattat tttttcaggg tcgtttttat gtgatttttt cccgtttcta
     7981 attttaaact aattgtttga tttttatatg attcgactct ttttaaatga attgttttgc
     8041 tttttttatt gagcatatat atattataca atttcaatca acaattgttg tgctttttaa
     8101 atgttgtttc tatttcattt ctagggtcga ttttattcga ttctagtttt aaaatcgtga
     8161 tttttaactt ctaaccttac tgagcagact gaatgaacat gtttcatcaa gattaacttc
     8221 attatagtta attattgaat cgtgctagtg gtttataaat cagctctatt taaattaatc
     8281 atcatcatta tttctattta atttttatta tttctttagg attattttta tgtgattttt
     8341 ttcacgttct agttttaaac tcaatgctca ttactgctca cccttgctga gcagtctttc
     8401 gttaagatta aattaattaa taaccattct agttttaaaa cagtgtctta aggcatagct
     8461 gatattttac aagtttacta tttaatatat ctcctttctt tttttagggt cgatttcatc
     8521 tgatttgatt aatttttatt atcttcacta agctttttgt tctaggtttt attctcagta
     8581 ttattactgt tatatttgaa ctgaatttct cctcatagtt tattcatttt atttatttat
     8641 tttttgttta ttcgctattt tgtattttag tttttaatta gaaattttat tagaatcgtt
     8701 ttgaactagt ttttatgcac tctaacatta atattaaatc ataactatta tttttattat
     8761 aatttatttg tgtttatctt tttgtgtagc tttcatgtgt ttttatttta gttttattat
     8821 gtttgcattg aattttttgg actagtttat ataacgatat acagaaaagc catgacaaca
     8881 aatgttttta taatacctat taagtataag ctaatcagat ttccactatt ctgttcaaag
     8941 ttaacattat tagccattct gatggatcat ttgacatttg tcatatgtct ttatgaacac
     9001 tgctctttca gatgccactt ttaaagctgt tttttacagt aaatgaacgt ttattattat
     9061 gacattattt cctaaggcct ctagagtaca gcaggatgtg gcgtgtgcat taccgtgtaa
     9121 ttatgctgac tgtaaatgtg catttataga ctgtcagcat gtctgactgt gcgctgttgt
     9181 gattggtcct gcacagacca ag
//
