LOCUS       sequence-220008-        9383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the 
            N-terminal sequence from zebrafish 1,4 galactosyltransferase 1 
            (GalT). Marks the Golgi complex..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9383)
  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 9383)
  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-GalT-mKate2
FEATURES             Location/Qualifiers
     source          1..9383
                     /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"
     protein_bind    complement(282..306)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             308..1003
                     /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"
     polyA_signal    1125..1206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1207..1227)
                     /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(1235..1251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1267..1401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1291..1310)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(1840..2027,2028..2039)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2073..2089)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2073..2089)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2126..2146)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2126..2144)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2165..2181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2178..2200)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2189..2205
                     /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(2213..2219,2220..2237,2238..2243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2258..2279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2396..2413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2567..3155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2647..2666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3326..4117,4118..4186))
                     /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     3949..3968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4187..4291)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4454..4475)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4824..4835,4836..4998)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5371..5387
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5407..5427
                     /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 catgccggac tccaccggga acttcagcct
       61 cctgcagcgg acctgctctc tggtggtgct gctgtgcttc ttacacatct ttgtcaccgt
      121 catttactac atgaggaact cggactctcg gccagctttc gcccagaacc agcagcagag
      181 accgacgata cacaggaaac tggcggagca gaggggcacc actgaggaca gcagacccgc
      241 ggccaacgcc tcgagcaacg gccaggagct gcagatctgc gacccagctt tcttgtacaa
      301 agtggctgtg agcgagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
      361 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
      421 ccagaccatg agaatcaagg cggtcgaggg cggccctctc cccttcgcct tcgacatcct
      481 ggctaccagc ttcatgtacg gcagcaaaac cttcatcaac cacacccagg gcatccccga
      541 cttctttaag cagtccttcc ccgagggctt cacatgggag agagtcacca catacgaaga
      601 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa
      661 cgtcaagatc agaggggtga acttcccatc caacggccct gtgatgcaga agaaaacact
      721 cggctgggag gcctccaccg agaccctgta ccccgctgac ggcggcctgg aaggcagagc
      781 cgacatggcc ctgaagctcg tgggcggggg ccacctgatc tgcaacttga agaccacata
      841 cagatccaag aaacccgcta agaacctcaa gatgcccggc gtctactatg tggacagaag
      901 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg aggtggctgt
      961 ggccagatac tgcgacctcc ctagcaaact ggggcacaga tgagcggccg cgactctaga
     1021 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1081 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     1141 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1201 cactgccaac tttattatac atagttgata attcactggc cgtcgtttta cggtaccatc
     1261 gatgatgatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1321 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1381 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1441 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     1501 tatgatcctc tagatcagat ctgcgaagat acggccacgg gtgctcttga tcctgtggct
     1561 gattttggac tgtgctgctc gcagctgctg atgaatcaca tacttcctcc attttcttcc
     1621 actgattgac tgttataatt tccctaattt ccaggtcaag gtgctgtgca ttgtggtaat
     1681 agatgtgaca tgacgtcact tccaaaggac caatgaacat gtctgaccaa tttcatataa
     1741 tgtgaaaacg attttcatag gcagaataaa taacatttaa attaaactgg gcatcagcgc
     1801 aattcaattg gtttggtaat agcaagggaa aatagaatga agtgatctcc aaaaaataag
     1861 tactttttga ctgtaaataa aattgtaagg agtaaaaagt actttttttt ctaaaaaaat
     1921 gtaattaagt aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag taaaaatccc
     1981 caaaaataat acttaagtac agtaatcaag taaaattact caagtacttt acacctctgg
     2041 ttcttgaccc cctaccttca gcaagcccag cagatccact agttctagag cggccgccac
     2101 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     2161 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2221 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2281 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2341 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2401 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2461 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2521 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2581 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2641 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2701 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2761 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2821 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2881 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2941 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3001 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3061 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3121 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3181 ggtctgacgc tcagtggaac gaaaactcac gttaaggcat tttggtcatg agattatcaa
     3241 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3301 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3361 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3421 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3481 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3541 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3601 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3661 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3721 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3781 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3841 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3901 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3961 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4021 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4081 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4141 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4201 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4261 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     4321 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4381 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4441 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4501 caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa ctcatcagcc
     4561 tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg gctactacat
     4621 ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca atgctcttgg
     4681 atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa tgccctctga
     4741 tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc acacaggtca
     4801 gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt
     4861 gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat ctggctagaa
     4921 tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt attaaggaag
     4981 taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa tctcaaaaaa
     5041 cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta tttaattatc
     5101 aaactataag acagacaatc taatgccagt acacgctact caaagttgta aaacctcaga
     5161 tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc tcttttgggg
     5221 ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc gggaagaggt
     5281 gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga ctccctggtg
     5341 tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat gattacgcca
     5401 agctatcaac tttgtataga aaagttggca cgcgcagatt ccgtgtgggc ctggtaatat
     5461 cttaccattt taagctattt taagattagg gaaaaaaatt aaataaaact actttaaaat
     5521 tactttcctt aaaaagcaac atatttattt agtttttgtt taactggctc aagattgtaa
     5581 tattttactt ttacatgttt ttttaagtac ctaaatgtgt tcaaaagtaa agtatgttat
     5641 tgtttttttt ttttaatgtt agctaagctt taaattgtaa tatattactt ttaaaagtga
     5701 ctttttaaaa atgtaaataa aaaagtaact ataaatgtgt aatgcatgtt atttctgtgg
     5761 gtttttaatt ttcatctcaa ctgggcttac aattttaaca taataaaaat caaatcaaat
     5821 caaatcaggc ccttatttgt cacatacact cttgtatact gaaataaact aaatgttgag
     5881 aaaaatttaa atacaccaaa ataacgcata ccttttttgt aattacctaa gtaaaacatt
     5941 tagttccttt tttaagtgac tcaagattgt aatctattaa ttgtaaaaga taaaactagt
     6001 gatcaatctg acgtgcttct atgcagcctg tgagattaat tgtgatgttt aattattaaa
     6061 caatttaaat ggatcaaaca tttcacatgt gcttattgtg tccaacaaaa ccagcagaag
     6121 cacttaaaaa tattgcatga caacatgagt ttttaatgta aattatatat acttcctaaa
     6181 cgaagttaat catatttcta aaataataat gtcattgaaa tatttaaata aatgataagc
     6241 tttagttgat tcaacataaa attgaaagca ataaggactt ttttttttta cagcggtgaa
     6301 gcaaaagcat gtaataatca tataatataa taaataaata aataaataat ggctcttggt
     6361 gcaaatatga acactttaaa taaatataga ataaatatga tcaataagtc ataacgatgt
     6421 gttttatgtt gattataatt taaacatgtt tctaacgcaa gttttaaaaa ctgttttaaa
     6481 gtattatttt aagttataag atcaatttga ctcaacattt tgtagtgaaa atatgttgtg
     6541 taaaattaat ataatataca ctataaaaat gcagggttcc acacaataca ttaatgttgt
     6601 cccaacacaa atcgattaag ttaacaaata tatgtggatt gaacataaaa aattaagttg
     6661 tcccaatgaa atctcaagaa ttgtgttgtt tcagctcatt ttaaataagt agttttaaca
     6721 agtaaaaaat aacatatttt tgagtgtgta tatatatata ttacaaaata taataatata
     6781 aaatgtaaaa gatataaacc atgctacata tatttaagag ctattttgca cattactttt
     6841 ttgcaagcca ttttagtgtt ttttttacaa atatatatta gctaaatata caatatctta
     6901 aatatacaat taatcagaca tgcaattaat gcatttaacc tacataaaaa tacatgatca
     6961 acatatgttt tatgctgatt acaactaaat caaacgtaat gaaattaatt tataaaagcg
     7021 atttaaatat gtcaatattt taagttaaaa tcacttgaaa cctactataa atgaacttaa
     7081 attaaaaaaa tctcctttaa ttccaatgca acagtgttgt tttttcagct gaggaccccc
     7141 ggtgcagatg gttggccccc ggcccgctat tctaaggaag cctttcttaa ttttagggcg
     7201 gcgctgcatg cgctcagatc agattcacgg gtggagaggg ttattttctc ttttttattt
     7261 tttcttgcct gctcatattt gggcctgtgt gaaccccagg ctccgcctcg gcccccagga
     7321 gccgctctta tttaaagggg atcctcgcct tcagtcttcg tcccagtgga agcatcagtc
     7381 ttgtgataac aacttggtga gtcctacaga ccagcgctcg actctaggct ttaaatgcat
     7441 tgatttctct ggattagcta ccgggagagt tttaactttg cttattagtc ttgtgctcga
     7501 actgctaggt ttaaagagtg ttttctctaa tgcatgcatg aagcgacagc tgaaaaccag
     7561 aaatagagcg tttgtcctcc tgttaaactc tagcctttct ttggcatttg tcagtccgag
     7621 tcttcagctg ccggcaggag attgggtttc agtaacactt gctgagcaca aactacacgt
     7681 gctttcttca gatttaattc attattaatc attctagatt taaatgatgt tcttactcat
     7741 tgctgattat tttagccttg cggagcagat taaatgagca cgtgtttcat taagattgat
     7801 ttaaatataa ttaatatttc accactgtag ttttaaaatt gccttctaag tcatggttga
     7861 tttttaatag gtatcggctc taattagatt actattttgg tttttaaatg ttatttctat
     7921 ttcatttgta ggctcgtttt taatgtgatt ttattagatt ctagttttaa aatcatctct
     7981 tagccttaat gagcagactg aatgaacatg tttcattaag attaacttca ttataatgaa
     8041 ttatttaatc attctagtgg gttataaatc agctctattt aaattaattg tcgtgtttat
     8101 ttctatttaa tttttgatta ttttttcagg gtcgttttta tgtgattttt tcccgtttct
     8161 aattttaaac taattgtttg atttttatat gattcgactc tttttaaatg aattgttttg
     8221 ctttttttat tgagcatata tatattatac aatttcaatc aacaattgtt gtgcttttta
     8281 aatgttgttt ctatttcatt tctagggtcg attttattcg attctagttt taaaatcgtg
     8341 atttttaact tctaacctta ctgagcagac tgaatgaaca tgtttcatca agattaactt
     8401 cattatagtt aattattgaa tcgtgctagt ggtttataaa tcagctctat ttaaattaat
     8461 catcatcatt atttctattt aatttttatt atttctttag gattattttt atgtgatttt
     8521 tttcacgttc tagttttaaa ctcaatgctc attactgctc acccttgctg agcagtcttt
     8581 cgttaagatt aaattaatta ataaccattc tagttttaaa acagtgtctt aaggcatagc
     8641 tgatatttta caagtttact atttaatata tctcctttct ttttttaggg tcgatttcat
     8701 ctgatttgat taatttttat tatcttcact aagctttttg ttctaggttt tattctcagt
     8761 attattactg ttatatttga actgaatttc tcctcatagt ttattcattt tatttattta
     8821 ttttttgttt attcgctatt ttgtatttta gtttttaatt agaaatttta ttagaatcgt
     8881 tttgaactag tttttatgca ctctaacatt aatattaaat cataactatt atttttatta
     8941 taatttattt gtgtttatct ttttgtgtag ctttcatgtg tttttatttt agttttatta
     9001 tgtttgcatt gaattttttg gactagttta tataacgata tacagaaaag ccatgacaac
     9061 aaatgttttt ataataccta ttaagtataa gctaatcaga tttccactat tctgttcaaa
     9121 gttaacatta ttagccattc tgatggatca tttgacattt gtcatatgtc tttatgaaca
     9181 ctgctctttc agatgccact tttaaagctg ttttttacag taaatgaacg tttattatta
     9241 tgacattatt tcctaaggcc tctagagtac agcaggatgt ggcgtgtgca ttaccgtgta
     9301 attatgctga ctgtaaatgt gcatttatag actgtcagca tgtctgactg tgcgctgttg
     9361 tgattggtcc tgcacagacc aag
//
