LOCUS Exported 9675 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to 3xPHD domain from Mouse inhibitor of growth family member 2 (ING2), codon optimised. Binds PtdIns5P. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-ING2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9675) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9675) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9675 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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(1499..1519) /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(1527..1543) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1559..1693 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1583..1602) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1637..1656 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2365..2381) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2365..2381) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2418..2438) /label=T3 /note="T3 promoter, forward primer" promoter complement(2418..2436) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2457..2473) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2470..2492) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2481..2497 /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(2505..2535) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2550..2571 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2688..2705) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2859..3447) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2939..2958) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3618..4478) /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 4241..4260 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4479..4583) /gene="bla" /label=AmpR promoter primer_bind complement(4746..4767) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5663..5679 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5699..5719 /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 ggtggcaagt ccagtagaat tcgcaattga 781 ccccaatgag ccaacttatt gtctttgcaa tcaagttagt tatggcgaga tgataggatg 841 cgacaacgag cagtgcccaa tagagtggtt ccatttcagt tgcgtatcat tgacatataa 901 accaaagggc aaatggtatt gcccaaagtg caggggtgac aatgagaaga cgatggacaa 961 gagtacagag aagacaaaaa aagagcgacg tgctcgtgag ttcgccagtc cggtggagtt 1021 cgcgattgac ccgaacgagc ccacgtactg cttgtgtaat caggtatcct acggggaaat 1081 gatcggatgc gacaatgaac agtgtcccat cgagtggttt cacttttcct gcgtgtcttt 1141 gacatataaa cctaaaggaa aatggtactg cccaaagtgt agaggagata atgaaaagac 1201 tatggataag tctaccgaga agactaaaaa ggaacgccga gcgagggagt tcgcaagtcc 1261 cgttgaattt gcgattgacc ctaatgagcc gacttactgc ctttgcaacc aggtttcata 1321 cggtgagatg atcggctgtg acaacgaaca atgcccaata gaatggttcc acttttcatg 1381 tgtgagtctt acatataaac ccaaggggaa atggtattgc cccaagtgtc gaggcgacaa 1441 tgaaaaaacg atggacaaga gcacagaaaa aaccaaaaaa gagagacgtg cgcggtaaca 1501 actttattat acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga 1561 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa 1621 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg 1681 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt 1741 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc 1801 tctagatcag atctgcgaag atacggccac gggtgctctt gatcctgtgg ctgattttgg 1861 actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt ccactgattg 1921 actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta atagatgtga 1981 catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat aatgtgaaaa 2041 cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc gcaattcaat 2101 tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt 2161 gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa 2221 gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata 2281 atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac 2341 cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc accgcggtgg 2401 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca 2461 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 2521 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 2581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 2641 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 2701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 2761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 2821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 2881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 2941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 3001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 3061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 3121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 3181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 3241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 3301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 3361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 3421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 3481 gctcagtgga acgaaaactc acgttaaggc attttggtca tgagattatc aaaaaggatc 3541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 3601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 3661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 3721 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 3781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 3841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 3901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 3961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 4021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 4081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 4141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 4201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 4261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 4321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 4381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 4441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 4501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 4561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg 4621 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 4681 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 4741 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 4801 gaaaaaccgt ctatcagggc gatggcccac gctggctaag aactcatcag cctccccggt 4861 ccatctaccc acgtaccaat gcaccaattg gccacaatga cggctactac atggtgccat 4921 tccttcctct ttataggaat ggagactacc tcctgtccaa caatgctctt ggatacgagt 4981 acgcctacct gttggaccca ggtcattgca caacaccaga aatgccctct gatctgcaaa 5041 agacgtgaat atctgttcag acacccatat ccactctgtt ccacacaggt cagaggtttg 5101 tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac 5161 aagtacttag ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt 5221 gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga gtattaagga agtaaaagta 5281 aaagcaagaa agaaaactag agattcttgt ttaagctttt aatctcaaaa aacattaaat 5341 gaaatgcata caaggtttta tcctgcttta gaactgtttg tatttaatta tcaaactata 5401 agacagacaa tctaatgcca gtacacgcta ctcaaagttg taaaacctca gatttaactt 5461 cagtagaagc tgattctcaa aattgttagt gtcaagccta gctcttttgg ggctgaaaag 5521 caatcctgca gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag gtgtattagt 5581 cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg tgtctgaaac 5641 acaggccaga tgggccctcg agcaggaaac agctatgacc atgattacgc caagctatca 5701 actttgtata gaaaagttgg cacgcgcaga ttccgtgtgg gcctggtaat atcttaccat 5761 tttaagctat tttaagatta gggaaaaaaa ttaaataaaa ctactttaaa attactttcc 5821 ttaaaaagca acatatttat ttagtttttg tttaactggc tcaagattgt aatattttac 5881 ttttacatgt ttttttaagt acctaaatgt gttcaaaagt aaagtatgtt attgtttttt 5941 ttttttaatg ttagctaagc tttaaattgt aatatattac ttttaaaagt gactttttaa 6001 aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg ttatttctgt gggtttttaa 6061 ttttcatctc aactgggctt acaattttaa cataataaaa atcaaatcaa atcaaatcag 6121 gcccttattt gtcacataca ctcttgtata ctgaaataaa ctaaatgttg agaaaaattt 6181 aaatacacca aaataacgca tacctttttt gtaattacct aagtaaaaca tttagttcct 6241 tttttaagtg actcaagatt gtaatctatt aattgtaaaa gataaaacta gtgatcaatc 6301 tgacgtgctt ctatgcagcc tgtgagatta attgtgatgt ttaattatta aacaatttaa 6361 atggatcaaa catttcacat gtgcttattg tgtccaacaa aaccagcaga agcacttaaa 6421 aatattgcat gacaacatga gtttttaatg taaattatat atacttccta aacgaagtta 6481 atcatatttc taaaataata atgtcattga aatatttaaa taaatgataa gctttagttg 6541 attcaacata aaattgaaag caataaggac tttttttttt tacagcggtg aagcaaaagc 6601 atgtaataat catataatat aataaataaa taaataaata atggctcttg gtgcaaatat 6661 gaacacttta aataaatata gaataaatat gatcaataag tcataacgat gtgttttatg 6721 ttgattataa tttaaacatg tttctaacgc aagttttaaa aactgtttta aagtattatt 6781 ttaagttata agatcaattt gactcaacat tttgtagtga aaatatgttg tgtaaaatta 6841 atataatata cactataaaa atgcagggtt ccacacaata cattaatgtt gtcccaacac 6901 aaatcgatta agttaacaaa tatatgtgga ttgaacataa aaaattaagt tgtcccaatg 6961 aaatctcaag aattgtgttg tttcagctca ttttaaataa gtagttttaa caagtaaaaa 7021 ataacatatt tttgagtgtg tatatatata tattacaaaa tataataata taaaatgtaa 7081 aagatataaa ccatgctaca tatatttaag agctattttg cacattactt ttttgcaagc 7141 cattttagtg ttttttttac aaatatatat tagctaaata tacaatatct taaatataca 7201 attaatcaga catgcaatta atgcatttaa cctacataaa aatacatgat caacatatgt 7261 tttatgctga ttacaactaa atcaaacgta atgaaattaa tttataaaag cgatttaaat 7321 atgtcaatat tttaagttaa aatcacttga aacctactat aaatgaactt aaattaaaaa 7381 aatctccttt aattccaatg caacagtgtt gttttttcag ctgaggaccc ccggtgcaga 7441 tggttggccc ccggcccgct attctaagga agcctttctt aattttaggg cggcgctgca 7501 tgcgctcaga tcagattcac gggtggagag ggttattttc tcttttttat tttttcttgc 7561 ctgctcatat ttgggcctgt gtgaacccca ggctccgcct cggcccccag gagccgctct 7621 tatttaaagg ggatcctcgc cttcagtctt cgtcccagtg gaagcatcag tcttgtgata 7681 acaacttggt gagtcctaca gaccagcgct cgactctagg ctttaaatgc attgatttct 7741 ctggattagc taccgggaga gttttaactt tgcttattag tcttgtgctc gaactgctag 7801 gtttaaagag tgttttctct aatgcatgca tgaagcgaca gctgaaaacc agaaatagag 7861 cgtttgtcct cctgttaaac tctagccttt ctttggcatt tgtcagtccg agtcttcagc 7921 tgccggcagg agattgggtt tcagtaacac ttgctgagca caaactacac gtgctttctt 7981 cagatttaat tcattattaa tcattctaga tttaaatgat gttcttactc attgctgatt 8041 attttagcct tgcggagcag attaaatgag cacgtgtttc attaagattg atttaaatat 8101 aattaatatt tcaccactgt agttttaaaa ttgccttcta agtcatggtt gatttttaat 8161 aggtatcggc tctaattaga ttactatttt ggtttttaaa tgttatttct atttcatttg 8221 taggctcgtt tttaatgtga ttttattaga ttctagtttt aaaatcatct cttagcctta 8281 atgagcagac tgaatgaaca tgtttcatta agattaactt cattataatg aattatttaa 8341 tcattctagt gggttataaa tcagctctat ttaaattaat tgtcgtgttt atttctattt 8401 aatttttgat tattttttca gggtcgtttt tatgtgattt tttcccgttt ctaattttaa 8461 actaattgtt tgatttttat atgattcgac tctttttaaa tgaattgttt tgcttttttt 8521 attgagcata tatatattat acaatttcaa tcaacaattg ttgtgctttt taaatgttgt 8581 ttctatttca tttctagggt cgattttatt cgattctagt tttaaaatcg tgatttttaa 8641 cttctaacct tactgagcag actgaatgaa catgtttcat caagattaac ttcattatag 8701 ttaattattg aatcgtgcta gtggtttata aatcagctct atttaaatta atcatcatca 8761 ttatttctat ttaattttta ttatttcttt aggattattt ttatgtgatt tttttcacgt 8821 tctagtttta aactcaatgc tcattactgc tcacccttgc tgagcagtct ttcgttaaga 8881 ttaaattaat taataaccat tctagtttta aaacagtgtc ttaaggcata gctgatattt 8941 tacaagttta ctatttaata tatctccttt ctttttttag ggtcgatttc atctgatttg 9001 attaattttt attatcttca ctaagctttt tgttctaggt tttattctca gtattattac 9061 tgttatattt gaactgaatt tctcctcata gtttattcat tttatttatt tattttttgt 9121 ttattcgcta ttttgtattt tagtttttaa ttagaaattt tattagaatc gttttgaact 9181 agtttttatg cactctaaca ttaatattaa atcataacta ttatttttat tataatttat 9241 ttgtgtttat ctttttgtgt agctttcatg tgtttttatt ttagttttat tatgtttgca 9301 ttgaattttt tggactagtt tatataacga tatacagaaa agccatgaca acaaatgttt 9361 ttataatacc tattaagtat aagctaatca gatttccact attctgttca aagttaacat 9421 tattagccat tctgatggat catttgacat ttgtcatatg tctttatgaa cactgctctt 9481 tcagatgcca cttttaaagc tgttttttac agtaaatgaa cgtttattat tatgacatta 9541 tttcctaagg cctctagagt acagcaggat gtggcgtgtg cattaccgtg taattatgct 9601 gactgtaaat gtgcatttat agactgtcag catgtctgac tgtgcgctgt tgtgattggt 9661 cctgcacaga ccaag //