LOCUS       sequence-206427-        9293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the 5' and 
            3' sequences from zebrafish folate receptor 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9293)
  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 9293)
  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-GPI-pA
FEATURES             Location/Qualifiers
     source          1..9293
                     /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             5189..5881
                     /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(5967..5991)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5993..6127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6190..6210)
                     /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(6218..6234)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6250..6384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(6823..7010,7011..7022)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7056..7072)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7056..7072)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7109..7129)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7109..7127)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7148..7164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7161..7183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7172..7188
                     /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(7196..7202,7203..7220,7221..7226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7241..7262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7379..7396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7550..8138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7630..7649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8309..9100,9101..9169))
                     /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     8932..8951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9170..9274)
                     /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 aanctttagt tgattcaaca taaaattgaa agcaataagg actttttttt
     1981 ttttacagcg gtgaagcaaa agcntgnaaa 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 ccatggacgc tttatttcac agacgtctgc tgctgctgct
     5161 gattgtactg ccggcctgca gcctcgaggt gagcgagctg attaaggaga acatgcacat
     5221 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
     5281 aggcaagccc tacgagggca cccagaccat gagaatcaag gcggtcgagg gcggccctct
     5341 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa
     5401 ccacacccag ggcatccccg acttctttaa gcagtccttc cccgagggct tcacatggga
     5461 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     5521 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc
     5581 tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagaccctgt accccgctga
     5641 cggcggcctg gaaggcagag ccgacatggc cctgaagctc gtgggcgggg gccacctgat
     5701 ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg
     5761 cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt
     5821 cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacag
     5881 aggatccctg aagggcggta cggagacggt cgccatggca acaggtctgc atctgatggc
     5941 gctgctggtg atgctgctgc gctgacaccc agctttcttg tacaaagtgg gggatccaga
     6001 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6061 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6121 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     6181 ggttttttcc aactttatta tacatagttg ataattcact ggccgtcgtt ttacggtacc
     6241 atcgatgatg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     6301 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     6361 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     6421 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     6481 gattatgatc ctctagatca gatctgcgaa gatacggcca cgggtgctct tgatcctgtg
     6541 gctgattttg gactgtgctg ctcgcagctg ctgatgaatc acatacttcc tccattttct
     6601 tccactgatt gactgttata atttccctaa tttccaggtc aaggtgctgt gcattgtggt
     6661 aatagatgtg acatgacgtc acttccaaag gaccaatgaa catgtctgac caatttcata
     6721 taatgtgaaa acgattttca taggcagaat aaataacatt taaattaaac tgggcatcag
     6781 cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat
     6841 aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa
     6901 aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat
     6961 ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc
     7021 tggttcttga ccccctacct tcagcaagcc cagcagatcc actagttcta gagcggccgc
     7081 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     7141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7981 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     8041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtg
//
