LOCUS Exported 9446 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to mouse 2x PH domain from HGF regulated tyrosine kinase substrate (Hgs). Binds PtdIns3P. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-2xFYVEhgs SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9446) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9446) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9446 /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(1270..1290) /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(1298..1314) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1330..1464 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1354..1373) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1408..1427 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2136..2152) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2136..2152) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2189..2209) /label=T3 /note="T3 promoter, forward primer" promoter complement(2189..2207) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2228..2244) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2241..2263) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2252..2268 /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(2276..2306) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2321..2342 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2459..2476) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2630..3218) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2710..2729) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3389..4249) /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 4012..4031 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4250..4354) /gene="bla" /label=AmpR promoter primer_bind complement(4517..4538) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5434..5450 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5470..5490 /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 ggctctcgag ctcaagcttc gaattgaatt 781 cgaaagtgat gccatgttcg ctgctgaaag agcccctgac tgggtggatg ctgaggaatg 841 ccatcggtgc agagtacagt ttggggtggt gacccgcaag catcactgcc gagcatgtgg 901 gcagatcttc tgtggcaagt gctcctccaa gtactccacc atccccaagt tcggcattga 961 gaaggaggtg cgcgtgtgtg agccctgcta tgagcagctg aacaagaagg cacaaggaca 1021 aggatccgaa agtgatgcca tgttcgctgc tgaaagagcc cctgactggg tggatgctga 1081 ggaatgccat cggtgcagag tacagtttgg ggtggtgacc cgcaagcatc actgccgagc 1141 atgtgggcag atcttctgtg gcaagtgctc ctccaagtac tccaccatcc ccaagttcgg 1201 cattgagaag gaggtgcgcg tgtgtgagcc ctgctatgag cagctgaaca agaaggcata 1261 agtcgacggc aactttatta tacatagttg ataattcact ggccgtcgtt ttacggtacc 1321 atcgatgatg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga 1381 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc 1441 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt 1501 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct 1561 gattatgatc ctctagatca gatctgcgaa gatacggcca cgggtgctct tgatcctgtg 1621 gctgattttg gactgtgctg ctcgcagctg ctgatgaatc acatacttcc tccattttct 1681 tccactgatt gactgttata atttccctaa tttccaggtc aaggtgctgt gcattgtggt 1741 aatagatgtg acatgacgtc acttccaaag gaccaatgaa catgtctgac caatttcata 1801 taatgtgaaa acgattttca taggcagaat aaataacatt taaattaaac tgggcatcag 1861 cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat 1921 aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa 1981 aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat 2041 ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc 2101 tggttcttga ccccctacct tcagcaagcc cagcagatcc actagttcta gagcggccgc 2161 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt 2221 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 2281 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 2341 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 2401 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 2461 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 2521 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 2581 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 2641 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 2701 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 2761 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 2821 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 2881 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 2941 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 3001 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 3061 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 3121 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 3181 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 3241 cggggtctga cgctcagtgg aacgaaaact cacgttaagg cattttggtc atgagattat 3301 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 3361 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 3421 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 3481 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 3541 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 3601 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 3661 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 3721 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 3781 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 3841 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 3901 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 3961 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 4021 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 4081 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 4141 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 4201 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 4261 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 4321 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta 4381 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 4441 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 4501 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 4561 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca cgctggctaa gaactcatca 4621 gcctccccgg tccatctacc cacgtaccaa tgcaccaatt ggccacaatg acggctacta 4681 catggtgcca ttccttcctc tttataggaa tggagactac ctcctgtcca acaatgctct 4741 tggatacgag tacgcctacc tgttggaccc aggtcattgc acaacaccag aaatgccctc 4801 tgatctgcaa aagacgtgaa tatctgttca gacacccata tccactctgt tccacacagg 4861 tcagaggttt gtccaggagt tcttgacaga ggtgtaaaaa gtactcaaaa attttactca 4921 agtgaaagta caagtactta gggaaaattt tactcaatta aaagtaaaag tatctggcta 4981 gaatcttact tgagtaaaag taaaaaagta ctccattaaa attgtacttg agtattaagg 5041 aagtaaaagt aaaagcaaga aagaaaacta gagattcttg tttaagcttt taatctcaaa 5101 aaacattaaa tgaaatgcat acaaggtttt atcctgcttt agaactgttt gtatttaatt 5161 atcaaactat aagacagaca atctaatgcc agtacacgct actcaaagtt gtaaaacctc 5221 agatttaact tcagtagaag ctgattctca aaattgttag tgtcaagcct agctcttttg 5281 gggctgaaaa gcaatcctgc agtgctgaaa agcctctcac aggcagccga tgcgggaaga 5341 ggtgtattag tcttgataga gaggctgcaa atagcaggaa acgtgagcag agactccctg 5401 gtgtctgaaa cacaggccag atgggccctc gagcaggaaa cagctatgac catgattacg 5461 ccaagctatc aactttgtat agaaaagttg gcacgcgcag attccgtgtg ggcctggtaa 5521 tatcttacca ttttaagcta ttttaagatt agggaaaaaa attaaataaa actactttaa 5581 aattactttc cttaaaaagc aacatattta tttagttttt gtttaactgg ctcaagattg 5641 taatatttta cttttacatg tttttttaag tacctaaatg tgttcaaaag taaagtatgt 5701 tattgttttt tttttttaat gttagctaag ctttaaattg taatatatta cttttaaaag 5761 tgacttttta aaaatgtaaa taaaaaagta actataaatg tgtaatgcat gttatttctg 5821 tgggttttta attttcatct caactgggct tacaatttta acataataaa aatcaaatca 5881 aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa actaaatgtt 5941 gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc taagtaaaac 6001 atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa agataaaact 6061 agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg tttaattatt 6121 aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca aaaccagcag 6181 aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata tatacttcct 6241 aaacgaagtt aatcatattt ctaaaataat aatgtcattg aaatatttaa ataaatgata 6301 agctttagtt gattcaacat aaaattgaaa gcaataagga cttttttttt ttacagcggt 6361 gaagcaaaag catgtaataa tcatataata taataaataa ataaataaat aatggctctt 6421 ggtgcaaata tgaacacttt aaataaatat agaataaata tgatcaataa gtcataacga 6481 tgtgttttat gttgattata atttaaacat gtttctaacg caagttttaa aaactgtttt 6541 aaagtattat tttaagttat aagatcaatt tgactcaaca ttttgtagtg aaaatatgtt 6601 gtgtaaaatt aatataatat acactataaa aatgcagggt tccacacaat acattaatgt 6661 tgtcccaaca caaatcgatt aagttaacaa atatatgtgg attgaacata aaaaattaag 6721 ttgtcccaat gaaatctcaa gaattgtgtt gtttcagctc attttaaata agtagtttta 6781 acaagtaaaa aataacatat ttttgagtgt gtatatatat atattacaaa atataataat 6841 ataaaatgta aaagatataa accatgctac atatatttaa gagctatttt gcacattact 6901 tttttgcaag ccattttagt gtttttttta caaatatata ttagctaaat atacaatatc 6961 ttaaatatac aattaatcag acatgcaatt aatgcattta acctacataa aaatacatga 7021 tcaacatatg ttttatgctg attacaacta aatcaaacgt aatgaaatta atttataaaa 7081 gcgatttaaa tatgtcaata ttttaagtta aaatcacttg aaacctacta taaatgaact 7141 taaattaaaa aaatctcctt taattccaat gcaacagtgt tgttttttca gctgaggacc 7201 cccggtgcag atggttggcc cccggcccgc tattctaagg aagcctttct taattttagg 7261 gcggcgctgc atgcgctcag atcagattca cgggtggaga gggttatttt ctctttttta 7321 ttttttcttg cctgctcata tttgggcctg tgtgaacccc aggctccgcc tcggccccca 7381 ggagccgctc ttatttaaag gggatcctcg ccttcagtct tcgtcccagt ggaagcatca 7441 gtcttgtgat aacaacttgg tgagtcctac agaccagcgc tcgactctag gctttaaatg 7501 cattgatttc tctggattag ctaccgggag agttttaact ttgcttatta gtcttgtgct 7561 cgaactgcta ggtttaaaga gtgttttctc taatgcatgc atgaagcgac agctgaaaac 7621 cagaaataga gcgtttgtcc tcctgttaaa ctctagcctt tctttggcat ttgtcagtcc 7681 gagtcttcag ctgccggcag gagattgggt ttcagtaaca cttgctgagc acaaactaca 7741 cgtgctttct tcagatttaa ttcattatta atcattctag atttaaatga tgttcttact 7801 cattgctgat tattttagcc ttgcggagca gattaaatga gcacgtgttt cattaagatt 7861 gatttaaata taattaatat ttcaccactg tagttttaaa attgccttct aagtcatggt 7921 tgatttttaa taggtatcgg ctctaattag attactattt tggtttttaa atgttatttc 7981 tatttcattt gtaggctcgt ttttaatgtg attttattag attctagttt taaaatcatc 8041 tcttagcctt aatgagcaga ctgaatgaac atgtttcatt aagattaact tcattataat 8101 gaattattta atcattctag tgggttataa atcagctcta tttaaattaa ttgtcgtgtt 8161 tatttctatt taatttttga ttattttttc agggtcgttt ttatgtgatt ttttcccgtt 8221 tctaatttta aactaattgt ttgattttta tatgattcga ctctttttaa atgaattgtt 8281 ttgctttttt tattgagcat atatatatta tacaatttca atcaacaatt gttgtgcttt 8341 ttaaatgttg tttctatttc atttctaggg tcgattttat tcgattctag ttttaaaatc 8401 gtgattttta acttctaacc ttactgagca gactgaatga acatgtttca tcaagattaa 8461 cttcattata gttaattatt gaatcgtgct agtggtttat aaatcagctc tatttaaatt 8521 aatcatcatc attatttcta tttaattttt attatttctt taggattatt tttatgtgat 8581 ttttttcacg ttctagtttt aaactcaatg ctcattactg ctcacccttg ctgagcagtc 8641 tttcgttaag attaaattaa ttaataacca ttctagtttt aaaacagtgt cttaaggcat 8701 agctgatatt ttacaagttt actatttaat atatctcctt tcttttttta gggtcgattt 8761 catctgattt gattaatttt tattatcttc actaagcttt ttgttctagg ttttattctc 8821 agtattatta ctgttatatt tgaactgaat ttctcctcat agtttattca ttttatttat 8881 ttattttttg tttattcgct attttgtatt ttagttttta attagaaatt ttattagaat 8941 cgttttgaac tagtttttat gcactctaac attaatatta aatcataact attattttta 9001 ttataattta tttgtgttta tctttttgtg tagctttcat gtgtttttat tttagtttta 9061 ttatgtttgc attgaatttt ttggactagt ttatataacg atatacagaa aagccatgac 9121 aacaaatgtt tttataatac ctattaagta taagctaatc agatttccac tattctgttc 9181 aaagttaaca ttattagcca ttctgatgga tcatttgaca tttgtcatat gtctttatga 9241 acactgctct ttcagatgcc acttttaaag ctgtttttta cagtaaatga acgtttatta 9301 ttatgacatt atttcctaag gcctctagag tacagcagga tgtggcgtgt gcattaccgt 9361 gtaattatgc tgactgtaaa tgtgcattta tagactgtca gcatgtctga ctgtgcgctg 9421 ttgtgattgg tcctgcacag accaag //