LOCUS       sequence-219966-        9408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to the 
            C-terminal domain from cow (Bos taurus) milk fat globue EGF-factor 8
            protein (MFGE8). Binds phosphatidylserine (PS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9408)
  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 9408)
  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-LactC2
FEATURES             Location/Qualifiers
     source          1..9408
                     /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(1232..1252)
                     /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(1260..1276)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1292..1426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1316..1335)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1370..1389
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1865..2052,2053..2064)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2098..2114)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2098..2114)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2151..2171)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2151..2169)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2190..2206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2203..2225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2214..2230
                     /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(2238..2244,2245..2262,2263..2268))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2283..2304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2421..2438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2592..3180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2672..2691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3351..4142,4143..4211))
                     /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     3974..3993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4212..4316)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4479..4500)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4849..4860,4861..5023)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5396..5412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5432..5452
                     /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 ggcttgcact gaacccctag gcctgaagga
      781 taataccatc cccaacaagc agatcacagc ctccagctac tacaaaacct ggggcctgag
      841 tgcctttagc tggtttccct actacgcacg actggataat cagggcaagt tcaacgcctg
      901 gaccgcccag accaacagtg cctctgagtg gctgcagatt gacctgggct cccagaagcg
      961 agtcacgggc atcatcaccc agggtgcccg agactttggc cacattcaat atgtggctgc
     1021 ctacagggtg gcctatggtg atgatggtgt gacctggact gagtacaagg acccgggggc
     1081 ctcagaaagc aagattttcc ctggtaacat ggacaataat tcccacaaga agaacatatt
     1141 tgagacgccg ttccaggctc gcttcgtgcg gatccagccc gtggcctggc acaaccgtat
     1201 caccctgcga gtggagctgc tgggctgtta gcaactttat tatacatagt tgataattca
     1261 ctggccgtcg ttttacggta ccatcgatga tgatccagac atgataagat acattgatga
     1321 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1381 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1441 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1501 cctctacaaa tgtggtatgg ctgattatga tcctctagat cagatctgcg aagatacggc
     1561 cacgggtgct cttgatcctg tggctgattt tggactgtgc tgctcgcagc tgctgatgaa
     1621 tcacatactt cctccatttt cttccactga ttgactgtta taatttccct aatttccagg
     1681 tcaaggtgct gtgcattgtg gtaatagatg tgacatgacg tcacttccaa aggaccaatg
     1741 aacatgtctg accaatttca tataatgtga aaacgatttt cataggcaga ataaataaca
     1801 tttaaattaa actgggcatc agcgcaattc aattggtttg gtaatagcaa gggaaaatag
     1861 aatgaagtga tctccaaaaa ataagtactt tttgactgta aataaaattg taaggagtaa
     1921 aaagtacttt tttttctaaa aaaatgtaat taagtaaaag taaaagtatt gatttttaat
     1981 tgtactcaag taaagtaaaa atccccaaaa ataatactta agtacagtaa tcaagtaaaa
     2041 ttactcaagt actttacacc tctggttctt gaccccctac cttcagcaag cccagcagat
     2101 ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     2161 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2221 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2281 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2341 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2401 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2461 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2521 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2581 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3001 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3061 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3121 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3181 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3241 ggcattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3301 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3361 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3421 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3481 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3541 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3601 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3661 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3721 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3781 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3841 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3901 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3961 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4021 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4081 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4141 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4201 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4321 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4381 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4441 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4501 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4561 cacgctggct aagaactcat cagcctcccc ggtccatcta cccacgtacc aatgcaccaa
     4621 ttggccacaa tgacggctac tacatggtgc cattccttcc tctttatagg aatggagact
     4681 acctcctgtc caacaatgct cttggatacg agtacgccta cctgttggac ccaggtcatt
     4741 gcacaacacc agaaatgccc tctgatctgc aaaagacgtg aatatctgtt cagacaccca
     4801 tatccactct gttccacaca ggtcagaggt ttgtccagga gttcttgaca gaggtgtaaa
     4861 aagtactcaa aaattttact caagtgaaag tacaagtact tagggaaaat tttactcaat
     4921 taaaagtaaa agtatctggc tagaatctta cttgagtaaa agtaaaaaag tactccatta
     4981 aaattgtact tgagtattaa ggaagtaaaa gtaaaagcaa gaaagaaaac tagagattct
     5041 tgtttaagct tttaatctca aaaaacatta aatgaaatgc atacaaggtt ttatcctgct
     5101 ttagaactgt ttgtatttaa ttatcaaact ataagacaga caatctaatg ccagtacacg
     5161 ctactcaaag ttgtaaaacc tcagatttaa cttcagtaga agctgattct caaaattgtt
     5221 agtgtcaagc ctagctcttt tggggctgaa aagcaatcct gcagtgctga aaagcctctc
     5281 acaggcagcc gatgcgggaa gaggtgtatt agtcttgata gagaggctgc aaatagcagg
     5341 aaacgtgagc agagactccc tggtgtctga aacacaggcc agatgggccc tcgagcagga
     5401 aacagctatg accatgatta cgccaagcta tcaactttgt atagaaaagt tggcacgcgc
     5461 agattccgtg tgggcctggt aatatcttac cattttaagc tattttaaga ttagggaaaa
     5521 aaattaaata aaactacttt aaaattactt tccttaaaaa gcaacatatt tatttagttt
     5581 ttgtttaact ggctcaagat tgtaatattt tacttttaca tgttttttta agtacctaaa
     5641 tgtgttcaaa agtaaagtat gttattgttt ttttttttta atgttagcta agctttaaat
     5701 tgtaatatat tacttttaaa agtgactttt taaaaatgta aataaaaaag taactataaa
     5761 tgtgtaatgc atgttatttc tgtgggtttt taattttcat ctcaactggg cttacaattt
     5821 taacataata aaaatcaaat caaatcaaat caggccctta tttgtcacat acactcttgt
     5881 atactgaaat aaactaaatg ttgagaaaaa tttaaataca ccaaaataac gcataccttt
     5941 tttgtaatta cctaagtaaa acatttagtt ccttttttaa gtgactcaag attgtaatct
     6001 attaattgta aaagataaaa ctagtgatca atctgacgtg cttctatgca gcctgtgaga
     6061 ttaattgtga tgtttaatta ttaaacaatt taaatggatc aaacatttca catgtgctta
     6121 ttgtgtccaa caaaaccagc agaagcactt aaaaatattg catgacaaca tgagttttta
     6181 atgtaaatta tatatacttc ctaaacgaag ttaatcatat ttctaaaata ataatgtcat
     6241 tgaaatattt aaataaatga taagctttag ttgattcaac ataaaattga aagcaataag
     6301 gacttttttt ttttacagcg gtgaagcaaa agcatgtaat aatcatataa tataataaat
     6361 aaataaataa ataatggctc ttggtgcaaa tatgaacact ttaaataaat atagaataaa
     6421 tatgatcaat aagtcataac gatgtgtttt atgttgatta taatttaaac atgtttctaa
     6481 cgcaagtttt aaaaactgtt ttaaagtatt attttaagtt ataagatcaa tttgactcaa
     6541 cattttgtag tgaaaatatg ttgtgtaaaa ttaatataat atacactata aaaatgcagg
     6601 gttccacaca atacattaat gttgtcccaa cacaaatcga ttaagttaac aaatatatgt
     6661 ggattgaaca taaaaaatta agttgtccca atgaaatctc aagaattgtg ttgtttcagc
     6721 tcattttaaa taagtagttt taacaagtaa aaaataacat atttttgagt gtgtatatat
     6781 atatattaca aaatataata atataaaatg taaaagatat aaaccatgct acatatattt
     6841 aagagctatt ttgcacatta cttttttgca agccatttta gtgttttttt tacaaatata
     6901 tattagctaa atatacaata tcttaaatat acaattaatc agacatgcaa ttaatgcatt
     6961 taacctacat aaaaatacat gatcaacata tgttttatgc tgattacaac taaatcaaac
     7021 gtaatgaaat taatttataa aagcgattta aatatgtcaa tattttaagt taaaatcact
     7081 tgaaacctac tataaatgaa cttaaattaa aaaaatctcc tttaattcca atgcaacagt
     7141 gttgtttttt cagctgagga cccccggtgc agatggttgg cccccggccc gctattctaa
     7201 ggaagccttt cttaatttta gggcggcgct gcatgcgctc agatcagatt cacgggtgga
     7261 gagggttatt ttctcttttt tattttttct tgcctgctca tatttgggcc tgtgtgaacc
     7321 ccaggctccg cctcggcccc caggagccgc tcttatttaa aggggatcct cgccttcagt
     7381 cttcgtccca gtggaagcat cagtcttgtg ataacaactt ggtgagtcct acagaccagc
     7441 gctcgactct aggctttaaa tgcattgatt tctctggatt agctaccggg agagttttaa
     7501 ctttgcttat tagtcttgtg ctcgaactgc taggtttaaa gagtgttttc tctaatgcat
     7561 gcatgaagcg acagctgaaa accagaaata gagcgtttgt cctcctgtta aactctagcc
     7621 tttctttggc atttgtcagt ccgagtcttc agctgccggc aggagattgg gtttcagtaa
     7681 cacttgctga gcacaaacta cacgtgcttt cttcagattt aattcattat taatcattct
     7741 agatttaaat gatgttctta ctcattgctg attattttag ccttgcggag cagattaaat
     7801 gagcacgtgt ttcattaaga ttgatttaaa tataattaat atttcaccac tgtagtttta
     7861 aaattgcctt ctaagtcatg gttgattttt aataggtatc ggctctaatt agattactat
     7921 tttggttttt aaatgttatt tctatttcat ttgtaggctc gtttttaatg tgattttatt
     7981 agattctagt tttaaaatca tctcttagcc ttaatgagca gactgaatga acatgtttca
     8041 ttaagattaa cttcattata atgaattatt taatcattct agtgggttat aaatcagctc
     8101 tatttaaatt aattgtcgtg tttatttcta tttaattttt gattattttt tcagggtcgt
     8161 ttttatgtga ttttttcccg tttctaattt taaactaatt gtttgatttt tatatgattc
     8221 gactcttttt aaatgaattg ttttgctttt tttattgagc atatatatat tatacaattt
     8281 caatcaacaa ttgttgtgct ttttaaatgt tgtttctatt tcatttctag ggtcgatttt
     8341 attcgattct agttttaaaa tcgtgatttt taacttctaa ccttactgag cagactgaat
     8401 gaacatgttt catcaagatt aacttcatta tagttaatta ttgaatcgtg ctagtggttt
     8461 ataaatcagc tctatttaaa ttaatcatca tcattatttc tatttaattt ttattatttc
     8521 tttaggatta tttttatgtg atttttttca cgttctagtt ttaaactcaa tgctcattac
     8581 tgctcaccct tgctgagcag tctttcgtta agattaaatt aattaataac cattctagtt
     8641 ttaaaacagt gtcttaaggc atagctgata ttttacaagt ttactattta atatatctcc
     8701 tttctttttt tagggtcgat ttcatctgat ttgattaatt tttattatct tcactaagct
     8761 ttttgttcta ggttttattc tcagtattat tactgttata tttgaactga atttctcctc
     8821 atagtttatt cattttattt atttattttt tgtttattcg ctattttgta ttttagtttt
     8881 taattagaaa ttttattaga atcgttttga actagttttt atgcactcta acattaatat
     8941 taaatcataa ctattatttt tattataatt tatttgtgtt tatctttttg tgtagctttc
     9001 atgtgttttt attttagttt tattatgttt gcattgaatt ttttggacta gtttatataa
     9061 cgatatacag aaaagccatg acaacaaatg tttttataat acctattaag tataagctaa
     9121 tcagatttcc actattctgt tcaaagttaa cattattagc cattctgatg gatcatttga
     9181 catttgtcat atgtctttat gaacactgct ctttcagatg ccacttttaa agctgttttt
     9241 tacagtaaat gaacgtttat tattatgaca ttatttccta aggcctctag agtacagcag
     9301 gatgtggcgt gtgcattacc gtgtaattat gctgactgta aatgtgcatt tatagactgt
     9361 cagcatgtct gactgtgcgc tgttgtgatt ggtcctgcac agaccaag
//
