LOCUS       sequence-220554-        9549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab1bb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9549)
  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 9549)
  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-rab1bb
FEATURES             Location/Qualifiers
     source          1..9549
                     /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(1373..1393)
                     /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(1401..1417)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1433..1567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1457..1476)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1511..1530
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(2006..2193,2194..2205)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2239..2255)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2239..2255)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2292..2312)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2292..2310)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2331..2347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2344..2366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2355..2371
                     /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(2379..2385,2386..2403,2404..2409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2424..2445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2562..2579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2733..3321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2813..2832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3492..4283,4284..4352))
                     /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     4115..4134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4353..4457)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4620..4641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4990..5001,5002..5164)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5537..5553
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5573..5593
                     /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 ggtgaaccct gaatatgact acttattcaa
      781 gcttcttctg attggagatt ctggagtggg aaaatcctgc attttgttgc gttttgctga
      841 cgacacctac acggagagtt tcatcagcac aattggagtg gattttaaga tccgaaccat
      901 tgagttggat ggaaaaacta ttaaactgca aatatgggac acggctggac aggagagatt
      961 ccgcactatt acatccagct actacagggg cgctcatggg atcatcgttg tatatgatgt
     1021 tacagatcag gaatcctaca acaatgtgaa acagtggcta aaggaaatcg atcgctatgc
     1081 gagtgaaaat gtcaacaaac tgcttgttgg aaataagtgt gacctgacta caaaaaaggt
     1141 tgtggactac actacagcta aggaatttgc tgactctctt ggcatccctt tccttgagac
     1201 gagtgccaaa aatgccacaa acgtggagca ggccttcatg accatggcag aggagataaa
     1261 gaagcgcatg agaccaggag cttcaggggg ctctgagaaa cctgacctta aaatcgaaag
     1321 caccccggtg caacagtcag gcggcggctg ctgttagaaa actcttgttt tccaacttta
     1381 ttatacatag ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga
     1441 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1501 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1561 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1621 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga
     1681 tcagatctgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg
     1741 ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt
     1801 ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac
     1861 gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt
     1921 tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt
     1981 ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt
     2041 aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa
     2101 gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt
     2161 aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta
     2221 ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc
     2281 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     2341 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2401 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2461 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2521 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2581 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2641 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2701 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2761 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2821 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2881 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2941 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3001 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3061 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3121 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3181 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3241 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3301 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3361 tggaacgaaa actcacgtta aggcattttg gtcatgagat tatcaaaaag gatcttcacc
     3421 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3481 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3541 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3601 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3661 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3721 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3781 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3841 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3901 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3961 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4021 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4081 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4141 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4201 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4261 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4321 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4381 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4441 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4501 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4561 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4621 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4681 ccgtctatca gggcgatggc ccacgctggc taagaactca tcagcctccc cggtccatct
     4741 acccacgtac caatgcacca attggccaca atgacggcta ctacatggtg ccattccttc
     4801 ctctttatag gaatggagac tacctcctgt ccaacaatgc tcttggatac gagtacgcct
     4861 acctgttgga cccaggtcat tgcacaacac cagaaatgcc ctctgatctg caaaagacgt
     4921 gaatatctgt tcagacaccc atatccactc tgttccacac aggtcagagg tttgtccagg
     4981 agttcttgac agaggtgtaa aaagtactca aaaattttac tcaagtgaaa gtacaagtac
     5041 ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa
     5101 aagtaaaaaa gtactccatt aaaattgtac ttgagtatta aggaagtaaa agtaaaagca
     5161 agaaagaaaa ctagagattc ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg
     5221 catacaaggt tttatcctgc tttagaactg tttgtattta attatcaaac tataagacag
     5281 acaatctaat gccagtacac gctactcaaa gttgtaaaac ctcagattta acttcagtag
     5341 aagctgattc tcaaaattgt tagtgtcaag cctagctctt ttggggctga aaagcaatcc
     5401 tgcagtgctg aaaagcctct cacaggcagc cgatgcggga agaggtgtat tagtcttgat
     5461 agagaggctg caaatagcag gaaacgtgag cagagactcc ctggtgtctg aaacacaggc
     5521 cagatgggcc ctcgagcagg aaacagctat gaccatgatt acgccaagct atcaactttg
     5581 tatagaaaag ttggcacgcg cagattccgt gtgggcctgg taatatctta ccattttaag
     5641 ctattttaag attagggaaa aaaattaaat aaaactactt taaaattact ttccttaaaa
     5701 agcaacatat ttatttagtt tttgtttaac tggctcaaga ttgtaatatt ttacttttac
     5761 atgttttttt aagtacctaa atgtgttcaa aagtaaagta tgttattgtt tttttttttt
     5821 aatgttagct aagctttaaa ttgtaatata ttacttttaa aagtgacttt ttaaaaatgt
     5881 aaataaaaaa gtaactataa atgtgtaatg catgttattt ctgtgggttt ttaattttca
     5941 tctcaactgg gcttacaatt ttaacataat aaaaatcaaa tcaaatcaaa tcaggccctt
     6001 atttgtcaca tacactcttg tatactgaaa taaactaaat gttgagaaaa atttaaatac
     6061 accaaaataa cgcatacctt ttttgtaatt acctaagtaa aacatttagt tcctttttta
     6121 agtgactcaa gattgtaatc tattaattgt aaaagataaa actagtgatc aatctgacgt
     6181 gcttctatgc agcctgtgag attaattgtg atgtttaatt attaaacaat ttaaatggat
     6241 caaacatttc acatgtgctt attgtgtcca acaaaaccag cagaagcact taaaaatatt
     6301 gcatgacaac atgagttttt aatgtaaatt atatatactt cctaaacgaa gttaatcata
     6361 tttctaaaat aataatgtca ttgaaatatt taaataaatg ataagcttta gttgattcaa
     6421 cataaaattg aaagcaataa ggactttttt tttttacagc ggtgaagcaa aagcatgtaa
     6481 taatcatata atataataaa taaataaata aataatggct cttggtgcaa atatgaacac
     6541 tttaaataaa tatagaataa atatgatcaa taagtcataa cgatgtgttt tatgttgatt
     6601 ataatttaaa catgtttcta acgcaagttt taaaaactgt tttaaagtat tattttaagt
     6661 tataagatca atttgactca acattttgta gtgaaaatat gttgtgtaaa attaatataa
     6721 tatacactat aaaaatgcag ggttccacac aatacattaa tgttgtccca acacaaatcg
     6781 attaagttaa caaatatatg tggattgaac ataaaaaatt aagttgtccc aatgaaatct
     6841 caagaattgt gttgtttcag ctcattttaa ataagtagtt ttaacaagta aaaaataaca
     6901 tatttttgag tgtgtatata tatatattac aaaatataat aatataaaat gtaaaagata
     6961 taaaccatgc tacatatatt taagagctat tttgcacatt acttttttgc aagccatttt
     7021 agtgtttttt ttacaaatat atattagcta aatatacaat atcttaaata tacaattaat
     7081 cagacatgca attaatgcat ttaacctaca taaaaataca tgatcaacat atgttttatg
     7141 ctgattacaa ctaaatcaaa cgtaatgaaa ttaatttata aaagcgattt aaatatgtca
     7201 atattttaag ttaaaatcac ttgaaaccta ctataaatga acttaaatta aaaaaatctc
     7261 ctttaattcc aatgcaacag tgttgttttt tcagctgagg acccccggtg cagatggttg
     7321 gcccccggcc cgctattcta aggaagcctt tcttaatttt agggcggcgc tgcatgcgct
     7381 cagatcagat tcacgggtgg agagggttat tttctctttt ttattttttc ttgcctgctc
     7441 atatttgggc ctgtgtgaac cccaggctcc gcctcggccc ccaggagccg ctcttattta
     7501 aaggggatcc tcgccttcag tcttcgtccc agtggaagca tcagtcttgt gataacaact
     7561 tggtgagtcc tacagaccag cgctcgactc taggctttaa atgcattgat ttctctggat
     7621 tagctaccgg gagagtttta actttgctta ttagtcttgt gctcgaactg ctaggtttaa
     7681 agagtgtttt ctctaatgca tgcatgaagc gacagctgaa aaccagaaat agagcgtttg
     7741 tcctcctgtt aaactctagc ctttctttgg catttgtcag tccgagtctt cagctgccgg
     7801 caggagattg ggtttcagta acacttgctg agcacaaact acacgtgctt tcttcagatt
     7861 taattcatta ttaatcattc tagatttaaa tgatgttctt actcattgct gattatttta
     7921 gccttgcgga gcagattaaa tgagcacgtg tttcattaag attgatttaa atataattaa
     7981 tatttcacca ctgtagtttt aaaattgcct tctaagtcat ggttgatttt taataggtat
     8041 cggctctaat tagattacta ttttggtttt taaatgttat ttctatttca tttgtaggct
     8101 cgtttttaat gtgattttat tagattctag ttttaaaatc atctcttagc cttaatgagc
     8161 agactgaatg aacatgtttc attaagatta acttcattat aatgaattat ttaatcattc
     8221 tagtgggtta taaatcagct ctatttaaat taattgtcgt gtttatttct atttaatttt
     8281 tgattatttt ttcagggtcg tttttatgtg attttttccc gtttctaatt ttaaactaat
     8341 tgtttgattt ttatatgatt cgactctttt taaatgaatt gttttgcttt ttttattgag
     8401 catatatata ttatacaatt tcaatcaaca attgttgtgc tttttaaatg ttgtttctat
     8461 ttcatttcta gggtcgattt tattcgattc tagttttaaa atcgtgattt ttaacttcta
     8521 accttactga gcagactgaa tgaacatgtt tcatcaagat taacttcatt atagttaatt
     8581 attgaatcgt gctagtggtt tataaatcag ctctatttaa attaatcatc atcattattt
     8641 ctatttaatt tttattattt ctttaggatt atttttatgt gatttttttc acgttctagt
     8701 tttaaactca atgctcatta ctgctcaccc ttgctgagca gtctttcgtt aagattaaat
     8761 taattaataa ccattctagt tttaaaacag tgtcttaagg catagctgat attttacaag
     8821 tttactattt aatatatctc ctttcttttt ttagggtcga tttcatctga tttgattaat
     8881 ttttattatc ttcactaagc tttttgttct aggttttatt ctcagtatta ttactgttat
     8941 atttgaactg aatttctcct catagtttat tcattttatt tatttatttt ttgtttattc
     9001 gctattttgt attttagttt ttaattagaa attttattag aatcgttttg aactagtttt
     9061 tatgcactct aacattaata ttaaatcata actattattt ttattataat ttatttgtgt
     9121 ttatcttttt gtgtagcttt catgtgtttt tattttagtt ttattatgtt tgcattgaat
     9181 tttttggact agtttatata acgatataca gaaaagccat gacaacaaat gtttttataa
     9241 tacctattaa gtataagcta atcagatttc cactattctg ttcaaagtta acattattag
     9301 ccattctgat ggatcatttg acatttgtca tatgtcttta tgaacactgc tctttcagat
     9361 gccactttta aagctgtttt ttacagtaaa tgaacgttta ttattatgac attatttcct
     9421 aaggcctcta gagtacagca ggatgtggcg tgtgcattac cgtgtaatta tgctgactgt
     9481 aaatgtgcat ttatagactg tcagcatgtc tgactgtgcg ctgttgtgat tggtcctgca
     9541 cagaccaag
//
