LOCUS Exported 9408 bp ds-DNA circular SYN 13-MAY-2021 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 actc1b-mKate2-LactC2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9408) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9408) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9408 /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(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" 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(2238..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(3351..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" 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 //