LOCUS       sequence-206430-        9203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the actin 
            binding lifeact sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9203)
  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 9203)
  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-Lifeact-mKate2
FEATURES             Location/Qualifiers
     source          1..9203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(514..525,526..688)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    5092..5116
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      5117..5126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5123..5173
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     protein_bind    complement(5175..5199)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5201..5896
                     /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    6018..6099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6100..6120)
                     /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(6128..6144)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6160..6294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6184..6203)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(6733..6920,6921..6932)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6966..6982)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6966..6982)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7019..7039)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7019..7037)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7058..7074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7071..7093)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7082..7098
                     /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(7106..7112,7113..7130,7131..7136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7151..7172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7289..7306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7460..8048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7540..7559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8219..9010,9011..9079))
                     /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     8842..8861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9080..9184)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttggca cgcgcagatt ccgtgtgggc
     1141 ctggtaatat cttaccattt taagctattt taagattagg gaaaaaaatt aaataaaact
     1201 actttaaaat tactttcctt aaaaagcaac atatttattt agtttttgtt taactggctc
     1261 aagattgtaa tattttactt ttacatgttt tttttaagta cctaaatgtg ttcaaaagta
     1321 aagtatgtta ttgttttttt ttttttaatg ttagctaagc tttaaattgt aatatattac
     1381 ttttaaaagt gactttttaa aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg
     1441 ttatttctgt gggtttttta attttcatct caactgggct tacaatttta acataataaa
     1501 aatcaaatca aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa
     1561 actaaatgtt gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc
     1621 taagtaaaac atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa
     1681 agataaaact agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg
     1741 tttaattatt aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca
     1801 aaaccagcag aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata
     1861 tatacttcct aaacgaagtt aatcatattt tctaaaataa taatgtcatt gaaatattta
     1921 aataaatgat aagctttagt tgattcaaca taaaattgaa agcaataagg actttttttt
     1981 ttttacagcg gtgaagcaaa agcatgtaaa taatcatata aatataataa ataaataaat
     2041 aaaataatgg ttcttggtgc aaatatgaac actttaaata aatatagaat aaatatgatc
     2101 aataagtcat aacgacgtgt tttatgttga ttataattta aacatgtttc taacgcaagt
     2161 tttaaaaact gttttaaagt attattttaa gttataagat caatttgact caacattttg
     2221 tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc aaggttccac
     2281 acaattcatt catgttgtcc caacacaaat cgattaagtt aacaaatata tgtggattga
     2341 acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc agctcatttt
     2401 aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata tatatatatt
     2461 tatatattac aaaatataat aatataaaat gtaaaagata taaaccatgc tacatatatt
     2521 taagagctat tttgcacatt acttttttgc aagccatttg agtgtttttt tacaaatata
     2581 tattagctaa atatacaata tcttaaatat acaattaatc agacatgcaa aaaatgcatt
     2641 taacctacat aaaaatacat gatcaacata tgttttatgc tgattacaac taaatcaaac
     2701 gtaatgaaat taatttataa aagcgattta aatatgtcaa tattttaagt taaaatcact
     2761 tgaaacctac tataaatgaa cttaaattta aaaaatctcc tttaattcca atgcagcact
     2821 gttgtttttt cagctgagga cccccggtgc agatggttgg cccccggccc gctattctaa
     2881 ggaagccttt cttaatttta gggcggcgct gcatgcgctc agatcagatt cacgggtgga
     2941 gagggttatt ttctcttttt tattttttct tgcctgctca tatttgggcc tgtgtgaacc
     3001 ccaggctccg cctcggcccc caggagccgc tcttatttaa aggggatcct cgccttcagt
     3061 cttcgtccca gtggaagcat cagtcttgtg ataacaactt ggtgagtcct acagaccagc
     3121 gctcgactct aggctttaaa tgcattgatt tctctggatt agctaccggg agagttttaa
     3181 ctttgcttat tagtcttgtg cttgaactgc taggtttaaa gagggttttc tctaatgcat
     3241 gcatgaagcg acagctgaaa accagaaata gagcgtttgt cctcctgtta aactctagcc
     3301 tttctttggc atttgtcagt ccgagtcttc agctgccggc aggagattgg gtttcagtaa
     3361 cacttgctga gcacaaacta cacgtgcttc cttcagattt aattcattat taatcattct
     3421 agatttaaat gatcttctta ctcattgctg attattttag ccttgcggag cagattaaat
     3481 gagcacgtgt ttattaagat tgatttaaat ataattaata tttcaccact gtagttttaa
     3541 aattgccttc taagtcatgg ttgattttta ataggtatcg gctctaatta gattactatt
     3601 ttggttttta aatgttattt ctatttcatt tgtaggctcg tttttaatgt gattttatta
     3661 gattctagtt ttaaaatcat cttttagcct tactgagcag actgaatgaa catgtttcat
     3721 taagattaac ttcattatag ttaattattt aatcattcta gtggattata aatcagctct
     3781 atttaaattg attgtcgtgt ttatttctat ttgatttttg attatttttt aaggtcgttt
     3841 ttatgtgatt tttccccgat tctaatttta aactaattgt ttgattttta tatgattcga
     3901 ctctttttaa atgaattgtt ttgctttttt tattgagcat atatatatat tatacaattt
     3961 caatcaacaa ttgttgtgct ttttaaatgt tgtttctatt tcatttctag ggtcgatttt
     4021 attcgattct agttttaaaa tcgtgatttc taactcctaa ccttactgag cagactgaat
     4081 gaacatgttt catcaagatt aacttcatta tagttaatta ttgaatcggg ctagtggttt
     4141 ataaatcagc tctattataa attaattgtc atcattattt ctatttaatt tttattattt
     4201 ttttaggatt atttttaagg gatttttttc acgttctagt tttaaactca atgctcatta
     4261 ctgctcaccc ttgctgagca gtctttcgtt aagattaaat taattaataa cccattctag
     4321 ttttaaaaca gtgtcttaag gcatagctga tattttacaa gtttactatt taatatatct
     4381 ccatttcttt ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa
     4441 gctttttgtt ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc
     4501 ctcatagttt attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt
     4561 ttttaattag aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa
     4621 tattaaatca taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct
     4681 ttcatgtgtt tttattttag ttttattatg tttgcattga attttttgga ctagtttata
     4741 taacgatata cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc
     4801 taatcagatt tccactattc tgttcaaagt taacattatt agccattctg atggatcatt
     4861 tgacatttgt catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt
     4921 ttttacagta aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag
     4981 caggatgtgg cgtgtgcatt actgtgtaat tatgctgact gtaaatgtgc atttatagac
     5041 tgtcagcatg tctgactgtg cgctgttgtg atttgtcctg cacagaccaa ggcaagtttg
     5101 tacaaaaaag caggctgcca ccatgggcgt ggccgacctg atcaagaagt tcgagagcat
     5161 cagcaaggag gagaacccag ctttcttgta caaagtggct gtgagcgagc tgattaagga
     5221 gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
     5281 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggcggtcga
     5341 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcaa
     5401 aaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct tccccgaggg
     5461 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     5521 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc
     5581 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggcctcca ccgagaccct
     5641 gtaccccgct gacggcggcc tggaaggcag agccgacatg gccctgaagc tcgtgggcgg
     5701 gggccacctg atctgcaact tgaagaccac atacagatcc aagaaacccg ctaagaacct
     5761 caagatgccc ggcgtctact atgtggacag aagactggaa agaatcaagg aggccgacaa
     5821 agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa
     5881 actggggcac agatgagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     5941 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     6001 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6061 gcatcacaaa tttcacaaat aaagcatttt tttcactgcc aactttatta tacatagttg
     6121 ataattcact ggccgtcgtt ttacggtacc atcgatgatg atccagacat gataagatac
     6181 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     6241 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     6301 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     6361 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctctagatca gatctgcgaa
     6421 gatacggcca cgggtgctct tgatcctgtg gctgattttg gactgtgctg ctcgcagctg
     6481 ctgatgaatc acatacttcc tccattttct tccactgatt gactgttata atttccctaa
     6541 tttccaggtc aaggtgctgt gcattgtggt aatagatgtg acatgacgtc acttccaaag
     6601 gaccaatgaa catgtctgac caatttcata taatgtgaaa acgattttca taggcagaat
     6661 aaataacatt taaattaaac tgggcatcag cgcaattcaa ttggtttggt aatagcaagg
     6721 gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa taaaattgta
     6781 aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta aaagtattga
     6841 tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag tacagtaatc
     6901 aagtaaaatt actcaagtac tttacacctc tggttcttga ccccctacct tcagcaagcc
     6961 cagcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
     7021 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     7081 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7141 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7201 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7261 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7321 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7381 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7441 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7501 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7561 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7621 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7681 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7741 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7801 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7861 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7921 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7981 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8041 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8101 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8161 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8221 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8281 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8341 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8401 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8461 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8521 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8581 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8641 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8701 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8761 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8821 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8881 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8941 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9001 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9061 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9121 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9181 cgcgcacatt tccccgaaaa gtg
//
