LOCUS Exported 9507 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to dominant negative Rac1. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-Rac1-DN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9507) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9507) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9507 /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(1331..1351) /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(1359..1375) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1391..1525 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1415..1434) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1469..1488 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2197..2213) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2197..2213) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2250..2270) /label=T3 /note="T3 promoter, forward primer" promoter complement(2250..2268) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2289..2305) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2302..2324) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2313..2329 /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(2337..2367) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2382..2403 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2520..2537) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2691..3279) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2771..2790) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3450..4310) /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 4073..4092 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4311..4415) /gene="bla" /label=AmpR promoter primer_bind complement(4578..4599) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5495..5511 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5531..5551 /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 ggctcaggcc ataaagtgtg tggtcgtcgg 781 ggacggggct gtgggaaaaa actgccttct gatcagctac accaccaatg cttttcctgg 841 ggaatacatt cccactgtgt ttgataatta ttctgcaaat gtgatggtcg atgggaaacc 901 agtaaacctg ggattgtggg atacagcagg acaagaggat tatgacaggc ttcgaccgct 961 ttcctaccct cagacggatg tgttcctgat ctgcttctct cttgtgagtc cggcctcatt 1021 tgaaaacgtc cgtgcaaagt ggtatccgga ggtcagacat cattgcccta acactccaat 1081 aattcttgtt gggactaagc ttgatttgcg agatgacaag gacaccatcg aaaagttaaa 1141 ggagaagaaa ctcactccca tcacctaccc tcaaggcctt gctatggcca aagaaatagg 1201 agctgtgaag tacctggagt gctctgcctt aacccaacgc ggccttaaaa ctgtgttcga 1261 tgaagccatc cgtgcagtgc tgtgtccacc tccggtgaag aggaggagga gaagatgtct 1321 ccttctgtga caactttatt atacatagtt gataattcac tggccgtcgt tttacggtac 1381 catcgatgat gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag 1441 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 1501 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 1561 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc 1621 tgattatgat cctctagatc agatctgcga agatacggcc acgggtgctc ttgatcctgt 1681 ggctgatttt ggactgtgct gctcgcagct gctgatgaat cacatacttc ctccattttc 1741 ttccactgat tgactgttat aatttcccta atttccaggt caaggtgctg tgcattgtgg 1801 taatagatgt gacatgacgt cacttccaaa ggaccaatga acatgtctga ccaatttcat 1861 ataatgtgaa aacgattttc ataggcagaa taaataacat ttaaattaaa ctgggcatca 1921 gcgcaattca attggtttgg taatagcaag ggaaaataga atgaagtgat ctccaaaaaa 1981 taagtacttt ttgactgtaa ataaaattgt aaggagtaaa aagtactttt ttttctaaaa 2041 aaatgtaatt aagtaaaagt aaaagtattg atttttaatt gtactcaagt aaagtaaaaa 2101 tccccaaaaa taatacttaa gtacagtaat caagtaaaat tactcaagta ctttacacct 2161 ctggttcttg accccctacc ttcagcaagc ccagcagatc cactagttct agagcggccg 2221 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg 2281 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 2341 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 2401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 2461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 2521 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 2581 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 2641 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 2701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 2761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2881 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 2941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 3001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 3061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 3121 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 3181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 3241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 3301 acggggtctg acgctcagtg gaacgaaaac tcacgttaag gcattttggt catgagatta 3361 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 3421 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 3481 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 3541 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 3601 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 3661 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 3721 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 3781 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 3841 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 3901 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 3961 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 4021 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 4081 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 4141 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 4201 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 4261 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 4321 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 4381 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 4441 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 4501 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 4561 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 4621 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc acgctggcta agaactcatc 4681 agcctccccg gtccatctac ccacgtacca atgcaccaat tggccacaat gacggctact 4741 acatggtgcc attccttcct ctttatagga atggagacta cctcctgtcc aacaatgctc 4801 ttggatacga gtacgcctac ctgttggacc caggtcattg cacaacacca gaaatgccct 4861 ctgatctgca aaagacgtga atatctgttc agacacccat atccactctg ttccacacag 4921 gtcagaggtt tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa aattttactc 4981 aagtgaaagt acaagtactt agggaaaatt ttactcaatt aaaagtaaaa gtatctggct 5041 agaatcttac ttgagtaaaa gtaaaaaagt actccattaa aattgtactt gagtattaag 5101 gaagtaaaag taaaagcaag aaagaaaact agagattctt gtttaagctt ttaatctcaa 5161 aaaacattaa atgaaatgca tacaaggttt tatcctgctt tagaactgtt tgtatttaat 5221 tatcaaacta taagacagac aatctaatgc cagtacacgc tactcaaagt tgtaaaacct 5281 cagatttaac ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc tagctctttt 5341 ggggctgaaa agcaatcctg cagtgctgaa aagcctctca caggcagccg atgcgggaag 5401 aggtgtatta gtcttgatag agaggctgca aatagcagga aacgtgagca gagactccct 5461 ggtgtctgaa acacaggcca gatgggccct cgagcaggaa acagctatga ccatgattac 5521 gccaagctat caactttgta tagaaaagtt ggcacgcgca gattccgtgt gggcctggta 5581 atatcttacc attttaagct attttaagat tagggaaaaa aattaaataa aactacttta 5641 aaattacttt ccttaaaaag caacatattt atttagtttt tgtttaactg gctcaagatt 5701 gtaatatttt acttttacat gtttttttaa gtacctaaat gtgttcaaaa gtaaagtatg 5761 ttattgtttt ttttttttaa tgttagctaa gctttaaatt gtaatatatt acttttaaaa 5821 gtgacttttt aaaaatgtaa ataaaaaagt aactataaat gtgtaatgca tgttatttct 5881 gtgggttttt aattttcatc tcaactgggc ttacaatttt aacataataa aaatcaaatc 5941 aaatcaaatc aggcccttat ttgtcacata cactcttgta tactgaaata aactaaatgt 6001 tgagaaaaat ttaaatacac caaaataacg catacctttt ttgtaattac ctaagtaaaa 6061 catttagttc cttttttaag tgactcaaga ttgtaatcta ttaattgtaa aagataaaac 6121 tagtgatcaa tctgacgtgc ttctatgcag cctgtgagat taattgtgat gtttaattat 6181 taaacaattt aaatggatca aacatttcac atgtgcttat tgtgtccaac aaaaccagca 6241 gaagcactta aaaatattgc atgacaacat gagtttttaa tgtaaattat atatacttcc 6301 taaacgaagt taatcatatt tctaaaataa taatgtcatt gaaatattta aataaatgat 6361 aagctttagt tgattcaaca taaaattgaa agcaataagg actttttttt tttacagcgg 6421 tgaagcaaaa gcatgtaata atcatataat ataataaata aataaataaa taatggctct 6481 tggtgcaaat atgaacactt taaataaata tagaataaat atgatcaata agtcataacg 6541 atgtgtttta tgttgattat aatttaaaca tgtttctaac gcaagtttta aaaactgttt 6601 taaagtatta ttttaagtta taagatcaat ttgactcaac attttgtagt gaaaatatgt 6661 tgtgtaaaat taatataata tacactataa aaatgcaggg ttccacacaa tacattaatg 6721 ttgtcccaac acaaatcgat taagttaaca aatatatgtg gattgaacat aaaaaattaa 6781 gttgtcccaa tgaaatctca agaattgtgt tgtttcagct cattttaaat aagtagtttt 6841 aacaagtaaa aaataacata tttttgagtg tgtatatata tatattacaa aatataataa 6901 tataaaatgt aaaagatata aaccatgcta catatattta agagctattt tgcacattac 6961 ttttttgcaa gccattttag tgtttttttt acaaatatat attagctaaa tatacaatat 7021 cttaaatata caattaatca gacatgcaat taatgcattt aacctacata aaaatacatg 7081 atcaacatat gttttatgct gattacaact aaatcaaacg taatgaaatt aatttataaa 7141 agcgatttaa atatgtcaat attttaagtt aaaatcactt gaaacctact ataaatgaac 7201 ttaaattaaa aaaatctcct ttaattccaa tgcaacagtg ttgttttttc agctgaggac 7261 ccccggtgca gatggttggc ccccggcccg ctattctaag gaagcctttc ttaattttag 7321 ggcggcgctg catgcgctca gatcagattc acgggtggag agggttattt tctctttttt 7381 attttttctt gcctgctcat atttgggcct gtgtgaaccc caggctccgc ctcggccccc 7441 aggagccgct cttatttaaa ggggatcctc gccttcagtc ttcgtcccag tggaagcatc 7501 agtcttgtga taacaacttg gtgagtccta cagaccagcg ctcgactcta ggctttaaat 7561 gcattgattt ctctggatta gctaccggga gagttttaac tttgcttatt agtcttgtgc 7621 tcgaactgct aggtttaaag agtgttttct ctaatgcatg catgaagcga cagctgaaaa 7681 ccagaaatag agcgtttgtc ctcctgttaa actctagcct ttctttggca tttgtcagtc 7741 cgagtcttca gctgccggca ggagattggg tttcagtaac acttgctgag cacaaactac 7801 acgtgctttc ttcagattta attcattatt aatcattcta gatttaaatg atgttcttac 7861 tcattgctga ttattttagc cttgcggagc agattaaatg agcacgtgtt tcattaagat 7921 tgatttaaat ataattaata tttcaccact gtagttttaa aattgccttc taagtcatgg 7981 ttgattttta ataggtatcg gctctaatta gattactatt ttggttttta aatgttattt 8041 ctatttcatt tgtaggctcg tttttaatgt gattttatta gattctagtt ttaaaatcat 8101 ctcttagcct taatgagcag actgaatgaa catgtttcat taagattaac ttcattataa 8161 tgaattattt aatcattcta gtgggttata aatcagctct atttaaatta attgtcgtgt 8221 ttatttctat ttaatttttg attatttttt cagggtcgtt tttatgtgat tttttcccgt 8281 ttctaatttt aaactaattg tttgattttt atatgattcg actcttttta aatgaattgt 8341 tttgcttttt ttattgagca tatatatatt atacaatttc aatcaacaat tgttgtgctt 8401 tttaaatgtt gtttctattt catttctagg gtcgatttta ttcgattcta gttttaaaat 8461 cgtgattttt aacttctaac cttactgagc agactgaatg aacatgtttc atcaagatta 8521 acttcattat agttaattat tgaatcgtgc tagtggttta taaatcagct ctatttaaat 8581 taatcatcat cattatttct atttaatttt tattatttct ttaggattat ttttatgtga 8641 tttttttcac gttctagttt taaactcaat gctcattact gctcaccctt gctgagcagt 8701 ctttcgttaa gattaaatta attaataacc attctagttt taaaacagtg tcttaaggca 8761 tagctgatat tttacaagtt tactatttaa tatatctcct ttcttttttt agggtcgatt 8821 tcatctgatt tgattaattt ttattatctt cactaagctt tttgttctag gttttattct 8881 cagtattatt actgttatat ttgaactgaa tttctcctca tagtttattc attttattta 8941 tttatttttt gtttattcgc tattttgtat tttagttttt aattagaaat tttattagaa 9001 tcgttttgaa ctagttttta tgcactctaa cattaatatt aaatcataac tattattttt 9061 attataattt atttgtgttt atctttttgt gtagctttca tgtgttttta ttttagtttt 9121 attatgtttg cattgaattt tttggactag tttatataac gatatacaga aaagccatga 9181 caacaaatgt ttttataata cctattaagt ataagctaat cagatttcca ctattctgtt 9241 caaagttaac attattagcc attctgatgg atcatttgac atttgtcata tgtctttatg 9301 aacactgctc tttcagatgc cacttttaaa gctgtttttt acagtaaatg aacgtttatt 9361 attatgacat tatttcctaa ggcctctaga gtacagcagg atgtggcgtg tgcattaccg 9421 tgtaattatg ctgactgtaa atgtgcattt atagactgtc agcatgtctg actgtgcgct 9481 gttgtgattg gtcctgcaca gaccaag //