LOCUS Exported 9522 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to dominant negative CDC42. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-CDC42-DN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9522) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9522) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9522 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(144..165) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1061..1077 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 1097..1117 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" protein_bind 5092..5116 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 5117..5126 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5123..5818 /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(5820..5844) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(6419..6439) /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(6447..6463) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 6479..6613 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6503..6522) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6557..6576 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(7285..7301) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7285..7301) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(7338..7358) /label=T3 /note="T3 promoter, forward primer" promoter complement(7338..7356) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7377..7393) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7390..7412) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7401..7417 /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(7425..7455) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7470..7491 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7608..7625) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7779..8367) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7859..7878) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8538..9398) /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 9161..9180 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9399..9503) /gene="bla" /label=AmpR promoter ORIGIN 1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa 241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg 301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca 361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa 421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc 481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt 541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat 601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt 661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa 721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta 781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta 841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc 901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc 961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga 1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat 1081 gattacgcca agctatcaac tttgtataga aaagttggca cgcgcagatt ccgtgtgggc 1141 ctggtaatat cttaccattt taagctattt taagattagg gaaaaaaatt aaataaaact 1201 actttaaaat tactttcctt aaaaagcaac atatttattt agtttttgtt taactggctc 1261 aagattgtaa tattttactt ttacatgttt tttttaagta cctaaatgtg ttcaaaagta 1321 aagtatgtta ttgttttttt ttttttaatg ttagctaagc tttaaattgt aatatattac 1381 ttttaaaagt gactttttaa aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg 1441 ttatttctgt gggtttttta attttcatct caactgggct tacaatttta acataataaa 1501 aatcaaatca aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa 1561 actaaatgtt gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc 1621 taagtaaaac atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa 1681 agataaaact agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg 1741 tttaattatt aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca 1801 aaaccagcag aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata 1861 tatacttcct aaacgaagtt aatcatattt tctaaaataa taatgtcatt gaaatattta 1921 aataaatgat aanctttagt tgattcaaca taaaattgaa agcaataagg actttttttt 1981 ttttacagcg gtgaagcaaa agcntgnaaa taatcatata aatataataa ataaataaat 2041 aaaataatgg ttcttggtgc aaatatgaac actttaaata aatatagaat aaatatgatc 2101 aataagtcat aacgacgtgt tttatgttga ttataattta aacatgtttc taacgcaagt 2161 tttaaaaact gttttaaagt attattttaa gttataagat caatttgact caacattttg 2221 tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc aaggttccac 2281 acaattcatt catgttgtcc caacacaaat cgattaagtt aacaaatata tgtggattga 2341 acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc agctcatttt 2401 aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata tatatatatt 2461 tatatattac aaaatataat aatataaaat gtaaaagata taaaccatgc tacatatatt 2521 taagagctat tttgcacatt acttttttgc aagccatttg agtgtttttt tacaaatata 2581 tattagctaa atatacaata tcttaaatat acaattaatc agacatgcaa aaaatgcatt 2641 taacctacat aaaaatacat gatcaacata tgttttatgc tgattacaac taaatcaaac 2701 gtaatgaaat taatttataa aagcgattta aatatgtcaa tattttaagt taaaatcact 2761 tgaaacctac tataaatgaa cttaaattta aaaaatctcc tttaattcca atgcagcact 2821 gttgtttttt cagctgagga cccccggtgc agatggttgg cccccggccc gctattctaa 2881 ggaagccttt cttaatttta gggcggcgct gcatgcgctc agatcagatt cacgggtgga 2941 gagggttatt ttctcttttt tattttttct tgcctgctca tatttgggcc tgtgtgaacc 3001 ccaggctccg cctcggcccc caggagccgc tcttatttaa aggggatcct cgccttcagt 3061 cttcgtccca gtggaagcat cagtcttgtg ataacaactt ggtgagtcct acagaccagc 3121 gctcgactct aggctttaaa tgcattgatt tctctggatt agctaccggg agagttttaa 3181 ctttgcttat tagtcttgtg cttgaactgc taggtttaaa gagggttttc tctaatgcat 3241 gcatgaagcg acagctgaaa accagaaata gagcgtttgt cctcctgtta aactctagcc 3301 tttctttggc atttgtcagt ccgagtcttc agctgccggc aggagattgg gtttcagtaa 3361 cacttgctga gcacaaacta cacgtgcttc cttcagattt aattcattat taatcattct 3421 agatttaaat gatcttctta ctcattgctg attattttag ccttgcggag cagattaaat 3481 gagcacgtgt ttattaagat tgatttaaat ataattaata tttcaccact gtagttttaa 3541 aattgccttc taagtcatgg ttgattttta ataggtatcg gctctaatta gattactatt 3601 ttggttttta aatgttattt ctatttcatt tgtaggctcg tttttaatgt gattttatta 3661 gattctagtt ttaaaatcat cttttagcct tactgagcag actgaatgaa catgtttcat 3721 taagattaac ttcattatag ttaattattt aatcattcta gtggattata aatcagctct 3781 atttaaattg attgtcgtgt ttatttctat ttgatttttg attatttttt aaggtcgttt 3841 ttatgtgatt tttccccgat tctaatttta aactaattgt ttgattttta tatgattcga 3901 ctctttttaa atgaattgtt ttgctttttt tattgagcat atatatatat tatacaattt 3961 caatcaacaa ttgttgtgct ttttaaatgt tgtttctatt tcatttctag ggtcgatttt 4021 attcgattct agttttaaaa tcgtgatttc taactcctaa ccttactgag cagactgaat 4081 gaacatgttt catcaagatt aacttcatta tagttaatta ttgaatcggg ctagtggttt 4141 ataaatcagc tctattataa attaattgtc atcattattt ctatttaatt tttattattt 4201 ttttaggatt atttttaagg gatttttttc acgttctagt tttaaactca atgctcatta 4261 ctgctcaccc ttgctgagca gtctttcgtt aagattaaat taattaataa cccattctag 4321 ttttaaaaca gtgtcttaag gcatagctga tattttacaa gtttactatt taatatatct 4381 ccatttcttt ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa 4441 gctttttgtt ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc 4501 ctcatagttt attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt 4561 ttttaattag aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa 4621 tattaaatca taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct 4681 ttcatgtgtt tttattttag ttttattatg tttgcattga attttttgga ctagtttata 4741 taacgatata cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc 4801 taatcagatt tccactattc tgttcaaagt taacattatt agccattctg atggatcatt 4861 tgacatttgt catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt 4921 ttttacagta aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag 4981 caggatgtgg cgtgtgcatt actgtgtaat tatgctgact gtaaatgtgc atttatagac 5041 tgtcagcatg tctgactgtg cgctgttgtg atttgtcctg cacagaccaa ggcaagtttg 5101 tacaaaaaag caggctgcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa 5161 gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg 5221 caagccctac gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc 5281 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca 5341 cacccagggc atccccgact tctttaagca gtccttcccc gagggcttca catgggagag 5401 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga 5461 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt 5521 gatgcagaag aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg 5581 cggcctggaa ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg 5641 caacttgaag accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt 5701 ctactatgtg gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga 5761 gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagaga 5821 cccagctttc ttgtacaaag tggctcagac gatcaagtgc gtcgttgttg gtgatggtgc 5881 agtgggtaaa aactgtctat taatctccta tacaactaat aaattcccct ccgaatatgt 5941 accaacggtc tttgataact atgcagttac ggttatgatt ggtggtgagc cgtacaccct 6001 ggggctgttt gatactgcag gtcaggaaga ttacgataga ttacgacccc tgagttaccc 6061 tcagacagat gtcttcttag tctgtttctc agttgtttcg ccttcttcat ttgaaaacgt 6121 caaagaaaag tgggtacccg agatcaccca ccactgtcca aagacccctt tcctgctggt 6181 ggggacgcag attgatctga gagatgatcc ttctactatc gaaaagcttg ccaagaacaa 6241 acagaaaccc atcactccag agacggcaga gaagctggcc cgcgatctga aggctgtcaa 6301 atacgtggaa tgctccgttc tgacgcagaa aggcctaaag aacgtatttg atgaggcgat 6361 tttggcagcc ctggagcccc cagagcccaa gaagaagcgt aaatgtgtgc tgctatgaca 6421 actttattat acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga 6481 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa 6541 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg 6601 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt 6661 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc 6721 tctagatcag atctgcgaag atacggccac gggtgctctt gatcctgtgg ctgattttgg 6781 actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt ccactgattg 6841 actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta atagatgtga 6901 catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat aatgtgaaaa 6961 cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc gcaattcaat 7021 tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt 7081 gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa 7141 gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata 7201 atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac 7261 cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc accgcggtgg 7321 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca 7381 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 7441 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 7501 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 7561 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 7621 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 7681 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 7741 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 7801 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 7861 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 7921 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 7981 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 8041 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 8101 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 8161 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 8221 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 8281 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 8341 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 8401 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 8461 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 8521 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 8581 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 8641 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 8701 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 8761 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 8821 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 8881 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 8941 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 9001 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 9061 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 9121 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 9181 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 9241 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 9301 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 9361 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 9421 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 9481 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tg //