LOCUS Exported 9614 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab5aa. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab5ab SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9614) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9614) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9614 /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(6511..6531) /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(6539..6555) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 6571..6705 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6595..6614) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6649..6668 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(7377..7393) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7377..7393) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(7430..7450) /label=T3 /note="T3 promoter, forward primer" promoter complement(7430..7448) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7469..7485) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7482..7504) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7493..7509 /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(7517..7547) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7562..7583 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7700..7717) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7871..8459) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7951..7970) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8630..9490) /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 9253..9272 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9491..9595) /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 tggtggcagg aagaggtgga gcaacgagac ctaacgggcc 5881 gaacaccggc aacaaaatct gtcagttcaa attagtgctg ctgggagagt ctgctgtggg 5941 gaagtcgagt ctagtgctgc gctttgtaaa aggacagttt catgagtttc aggaaagcac 6001 aataggagcc gccttcttga ctcaaacatt atgtctggat gacacaacgg cgaagtttga 6061 gatctgggat acagctggcc aggagcgcta ccacagtctg gccccaatgt attacagagg 6121 agcacaggcg gccatagtgg tgtatgacat cacaaatgaa gaatcatttg cacgtgcgaa 6181 aaactgggtg aaagagcttc aaagacaagc cagtccaaat attgtaatag ctttggctgg 6241 gaacaaggca gaccttgcca acaagagagc gttggacttt caggatgcgc aatcatatgc 6301 agatgacaac agtttgctct ttatggaaac gtcggcaaag acttcaatga acgtcagtga 6361 gatttttatg gcgatcgcaa agaagctccc gaagaacgag ccccagcccg caggtgccaa 6421 cacagcccgc aacagaggag tggatctaac agaaacagca cagcccgcca aaggcccctg 6481 ctgtagcacc taaaccacgc tcctcacctc caactttatt atacatagtt gataattcac 6541 tggccgtcgt tttacggtac catcgatgat gatccagaca tgataagata cattgatgag 6601 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat 6661 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc 6721 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac 6781 ctctacaaat gtggtatggc tgattatgat cctctagatc agatctgcga agatacggcc 6841 acgggtgctc ttgatcctgt ggctgatttt ggactgtgct gctcgcagct gctgatgaat 6901 cacatacttc ctccattttc ttccactgat tgactgttat aatttcccta atttccaggt 6961 caaggtgctg tgcattgtgg taatagatgt gacatgacgt cacttccaaa ggaccaatga 7021 acatgtctga ccaatttcat ataatgtgaa aacgattttc ataggcagaa taaataacat 7081 ttaaattaaa ctgggcatca gcgcaattca attggtttgg taatagcaag ggaaaataga 7141 atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt aaggagtaaa 7201 aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg atttttaatt 7261 gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat caagtaaaat 7321 tactcaagta ctttacacct ctggttcttg accccctacc ttcagcaagc ccagcagatc 7381 cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag 7441 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 7501 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 7561 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 7621 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 7681 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 7741 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 7801 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 7861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 7921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 7981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 8041 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 8101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 8161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 8221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 8281 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 8341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 8401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 8461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 8521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 8581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 8641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 8701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 8761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 8821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 8881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 8941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 9001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 9061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 9121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 9181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 9241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 9301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 9361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 9421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 9481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 9541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 9601 ttccccgaaa agtg //