LOCUS       sequence-221216-        9519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab22a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9519)
  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 9519)
  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-rab22a
FEATURES             Location/Qualifiers
     source          1..9519
                     /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(1343..1363)
                     /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(1371..1387)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1403..1537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1427..1446)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1481..1500
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1976..2163,2164..2175)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2209..2225)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2209..2225)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2262..2282)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2262..2280)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2301..2317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2314..2336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2325..2341
                     /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(2349..2355,2356..2373,2374..2379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2394..2415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2532..2549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2703..3291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2783..2802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3462..4253,4254..4322))
                     /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     4085..4104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4323..4427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4590..4611)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4960..4971,4972..5134)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5507..5523
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5543..5563
                     /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 ggctacgcta agagagctga aagtatgcct
      781 tcttggggac acaggagttg gaaagtcaag cattgtgtgt agatttgtgg aggatagttt
      841 tgatccaaac ataaatccaa caattggagc atcgtttatg acaaagactg tgcaatacca
      901 aaacgagctt cacaaattcc tcatctggga cacagcagga caagaaaggt ttcgtgctct
      961 ggctccgatg tactacagag gctctgctgc ggccatcatc gtctatgaca tcactaaaga
     1021 ggaatcattc cagacattga agaactgggt gaaggagctt cgccagcacg gaccacccaa
     1081 catagtggtg gctatcgctg gcaacaagtg cgatctctcc gatgccagag aggtatcaga
     1141 gaaggacgcc aaagactacg ctgactctat tcatgctata tttatcgaaa ccagtgccaa
     1201 aaatgccatt aacatcaatg aggtcttcac acagataagt gaaaggatcc cggttttaga
     1261 tgcggagggc gggtctgcag tcaagggttt taaactccgt cgccaaccat cagtatcgac
     1321 cagagaacgc acttgctgct gacaacttta ttatacatag ttgataattc actggccgtc
     1381 gttttacggt accatcgatg atgatccaga catgataaga tacattgatg agtttggaca
     1441 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1501 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1561 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1621 atgtggtatg gctgattatg atcctctaga tcagatctgc gaagatacgg ccacgggtgc
     1681 tcttgatcct gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact
     1741 tcctccattt tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc
     1801 tgtgcattgt ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct
     1861 gaccaatttc atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta
     1921 aactgggcat cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg
     1981 atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt
     2041 ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa
     2101 gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag
     2161 tactttacac ctctggttct tgacccccta ccttcagcaa gcccagcaga tccactagtt
     2221 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     2281 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2341 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2401 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2461 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2521 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2581 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2641 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2701 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2761 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2821 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2881 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2941 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3001 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3061 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3121 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3181 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3241 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3301 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta aggcattttg
     3361 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3421 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3481 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3541 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3601 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3661 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3721 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3781 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3841 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3901 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3961 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4021 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4081 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4141 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4201 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4261 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4321 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4381 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4441 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4501 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4561 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4621 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccacgctggc
     4681 taagaactca tcagcctccc cggtccatct acccacgtac caatgcacca attggccaca
     4741 atgacggcta ctacatggtg ccattccttc ctctttatag gaatggagac tacctcctgt
     4801 ccaacaatgc tcttggatac gagtacgcct acctgttgga cccaggtcat tgcacaacac
     4861 cagaaatgcc ctctgatctg caaaagacgt gaatatctgt tcagacaccc atatccactc
     4921 tgttccacac aggtcagagg tttgtccagg agttcttgac agaggtgtaa aaagtactca
     4981 aaaattttac tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa ttaaaagtaa
     5041 aagtatctgg ctagaatctt acttgagtaa aagtaaaaaa gtactccatt aaaattgtac
     5101 ttgagtatta aggaagtaaa agtaaaagca agaaagaaaa ctagagattc ttgtttaagc
     5161 ttttaatctc aaaaaacatt aaatgaaatg catacaaggt tttatcctgc tttagaactg
     5221 tttgtattta attatcaaac tataagacag acaatctaat gccagtacac gctactcaaa
     5281 gttgtaaaac ctcagattta acttcagtag aagctgattc tcaaaattgt tagtgtcaag
     5341 cctagctctt ttggggctga aaagcaatcc tgcagtgctg aaaagcctct cacaggcagc
     5401 cgatgcggga agaggtgtat tagtcttgat agagaggctg caaatagcag gaaacgtgag
     5461 cagagactcc ctggtgtctg aaacacaggc cagatgggcc ctcgagcagg aaacagctat
     5521 gaccatgatt acgccaagct atcaactttg tatagaaaag ttggcacgcg cagattccgt
     5581 gtgggcctgg taatatctta ccattttaag ctattttaag attagggaaa aaaattaaat
     5641 aaaactactt taaaattact ttccttaaaa agcaacatat ttatttagtt tttgtttaac
     5701 tggctcaaga ttgtaatatt ttacttttac atgttttttt aagtacctaa atgtgttcaa
     5761 aagtaaagta tgttattgtt tttttttttt aatgttagct aagctttaaa ttgtaatata
     5821 ttacttttaa aagtgacttt ttaaaaatgt aaataaaaaa gtaactataa atgtgtaatg
     5881 catgttattt ctgtgggttt ttaattttca tctcaactgg gcttacaatt ttaacataat
     5941 aaaaatcaaa tcaaatcaaa tcaggccctt atttgtcaca tacactcttg tatactgaaa
     6001 taaactaaat gttgagaaaa atttaaatac accaaaataa cgcatacctt ttttgtaatt
     6061 acctaagtaa aacatttagt tcctttttta agtgactcaa gattgtaatc tattaattgt
     6121 aaaagataaa actagtgatc aatctgacgt gcttctatgc agcctgtgag attaattgtg
     6181 atgtttaatt attaaacaat ttaaatggat caaacatttc acatgtgctt attgtgtcca
     6241 acaaaaccag cagaagcact taaaaatatt gcatgacaac atgagttttt aatgtaaatt
     6301 atatatactt cctaaacgaa gttaatcata tttctaaaat aataatgtca ttgaaatatt
     6361 taaataaatg ataagcttta gttgattcaa cataaaattg aaagcaataa ggactttttt
     6421 tttttacagc ggtgaagcaa aagcatgtaa taatcatata atataataaa taaataaata
     6481 aataatggct cttggtgcaa atatgaacac tttaaataaa tatagaataa atatgatcaa
     6541 taagtcataa cgatgtgttt tatgttgatt ataatttaaa catgtttcta acgcaagttt
     6601 taaaaactgt tttaaagtat tattttaagt tataagatca atttgactca acattttgta
     6661 gtgaaaatat gttgtgtaaa attaatataa tatacactat aaaaatgcag ggttccacac
     6721 aatacattaa tgttgtccca acacaaatcg attaagttaa caaatatatg tggattgaac
     6781 ataaaaaatt aagttgtccc aatgaaatct caagaattgt gttgtttcag ctcattttaa
     6841 ataagtagtt ttaacaagta aaaaataaca tatttttgag tgtgtatata tatatattac
     6901 aaaatataat aatataaaat gtaaaagata taaaccatgc tacatatatt taagagctat
     6961 tttgcacatt acttttttgc aagccatttt agtgtttttt ttacaaatat atattagcta
     7021 aatatacaat atcttaaata tacaattaat cagacatgca attaatgcat ttaacctaca
     7081 taaaaataca tgatcaacat atgttttatg ctgattacaa ctaaatcaaa cgtaatgaaa
     7141 ttaatttata aaagcgattt aaatatgtca atattttaag ttaaaatcac ttgaaaccta
     7201 ctataaatga acttaaatta aaaaaatctc ctttaattcc aatgcaacag tgttgttttt
     7261 tcagctgagg acccccggtg cagatggttg gcccccggcc cgctattcta aggaagcctt
     7321 tcttaatttt agggcggcgc tgcatgcgct cagatcagat tcacgggtgg agagggttat
     7381 tttctctttt ttattttttc ttgcctgctc atatttgggc ctgtgtgaac cccaggctcc
     7441 gcctcggccc ccaggagccg ctcttattta aaggggatcc tcgccttcag tcttcgtccc
     7501 agtggaagca tcagtcttgt gataacaact tggtgagtcc tacagaccag cgctcgactc
     7561 taggctttaa atgcattgat ttctctggat tagctaccgg gagagtttta actttgctta
     7621 ttagtcttgt gctcgaactg ctaggtttaa agagtgtttt ctctaatgca tgcatgaagc
     7681 gacagctgaa aaccagaaat agagcgtttg tcctcctgtt aaactctagc ctttctttgg
     7741 catttgtcag tccgagtctt cagctgccgg caggagattg ggtttcagta acacttgctg
     7801 agcacaaact acacgtgctt tcttcagatt taattcatta ttaatcattc tagatttaaa
     7861 tgatgttctt actcattgct gattatttta gccttgcgga gcagattaaa tgagcacgtg
     7921 tttcattaag attgatttaa atataattaa tatttcacca ctgtagtttt aaaattgcct
     7981 tctaagtcat ggttgatttt taataggtat cggctctaat tagattacta ttttggtttt
     8041 taaatgttat ttctatttca tttgtaggct cgtttttaat gtgattttat tagattctag
     8101 ttttaaaatc atctcttagc cttaatgagc agactgaatg aacatgtttc attaagatta
     8161 acttcattat aatgaattat ttaatcattc tagtgggtta taaatcagct ctatttaaat
     8221 taattgtcgt gtttatttct atttaatttt tgattatttt ttcagggtcg tttttatgtg
     8281 attttttccc gtttctaatt ttaaactaat tgtttgattt ttatatgatt cgactctttt
     8341 taaatgaatt gttttgcttt ttttattgag catatatata ttatacaatt tcaatcaaca
     8401 attgttgtgc tttttaaatg ttgtttctat ttcatttcta gggtcgattt tattcgattc
     8461 tagttttaaa atcgtgattt ttaacttcta accttactga gcagactgaa tgaacatgtt
     8521 tcatcaagat taacttcatt atagttaatt attgaatcgt gctagtggtt tataaatcag
     8581 ctctatttaa attaatcatc atcattattt ctatttaatt tttattattt ctttaggatt
     8641 atttttatgt gatttttttc acgttctagt tttaaactca atgctcatta ctgctcaccc
     8701 ttgctgagca gtctttcgtt aagattaaat taattaataa ccattctagt tttaaaacag
     8761 tgtcttaagg catagctgat attttacaag tttactattt aatatatctc ctttcttttt
     8821 ttagggtcga tttcatctga tttgattaat ttttattatc ttcactaagc tttttgttct
     8881 aggttttatt ctcagtatta ttactgttat atttgaactg aatttctcct catagtttat
     8941 tcattttatt tatttatttt ttgtttattc gctattttgt attttagttt ttaattagaa
     9001 attttattag aatcgttttg aactagtttt tatgcactct aacattaata ttaaatcata
     9061 actattattt ttattataat ttatttgtgt ttatcttttt gtgtagcttt catgtgtttt
     9121 tattttagtt ttattatgtt tgcattgaat tttttggact agtttatata acgatataca
     9181 gaaaagccat gacaacaaat gtttttataa tacctattaa gtataagcta atcagatttc
     9241 cactattctg ttcaaagtta acattattag ccattctgat ggatcatttg acatttgtca
     9301 tatgtcttta tgaacactgc tctttcagat gccactttta aagctgtttt ttacagtaaa
     9361 tgaacgttta ttattatgac attatttcct aaggcctcta gagtacagca ggatgtggcg
     9421 tgtgcattac cgtgtaatta tgctgactgt aaatgtgcat ttatagactg tcagcatgtc
     9481 tgactgtgcg ctgttgtgat tggtcctgca cagaccaag
//
