LOCUS Exported 9555 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab6ba. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab6bb SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9555) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9555) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9555 /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(1379..1399) /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(1407..1423) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1439..1573 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1463..1482) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1517..1536 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2245..2261) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2245..2261) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2298..2318) /label=T3 /note="T3 promoter, forward primer" promoter complement(2298..2316) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2337..2353) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2350..2372) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2361..2377 /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(2385..2415) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2430..2451 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2568..2585) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2739..3327) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2819..2838) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3498..4358) /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 4121..4140 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4359..4463) /gene="bla" /label=AmpR promoter primer_bind complement(4626..4647) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5543..5559 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5579..5599 /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 ggtgtccatg ggaagtgact ttgggaaccc 781 tttacggaaa ttcaaacttg tttttttagg ggagcaaagc gttgggaaga catcgctgat 841 aacaaggttc atgtatgaca gttttgacaa cacttatcag gcaactattg gcattgactt 901 tttatcaaaa accatgtatc tggaggaccg aactgtgagg ttacagctat gggacactgc 961 agggcaggag cgttttagga gtcttatccc gagctacatt agagactcca cagtggctgt 1021 ggtggtgtat gatatcacaa atatcaattc attccagcta acctctaaat ggattgatga 1081 tgtcagaaca gagaggggaa gtgatgtcat tatcatgctg gtgggcaata aaacagatct 1141 agaggaaaag aggcaaatca ctatagagga gggggagcag cgggccaaag agttgaatgt 1201 gatgtttatc gagactagtg caaagaccgg cagtaatgtt aaacagttgt ttcggcgagt 1261 ggccgctgct ttgcctggta tggagagctt ggatgataat aacaaagaag gaatgattga 1321 cattaagctg gacaaacagc cggatcctcc ggccactgaa agcggctgct cctgttagca 1381 actttattat acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga 1441 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa 1501 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg 1561 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt 1621 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc 1681 tctagatcag atctgcgaag atacggccac gggtgctctt gatcctgtgg ctgattttgg 1741 actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt ccactgattg 1801 actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta atagatgtga 1861 catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat aatgtgaaaa 1921 cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc gcaattcaat 1981 tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt 2041 gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa 2101 gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata 2161 atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct ggttcttgac 2221 cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc accgcggtgg 2281 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca 2341 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 2401 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 2461 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 2521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 2581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 2641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 2701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 2761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 2821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 2881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 2941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 3001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 3061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 3121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 3181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 3241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 3301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 3361 gctcagtgga acgaaaactc acgttaaggc attttggtca tgagattatc aaaaaggatc 3421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 3481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 3541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 3601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 3661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 3721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 3781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 3841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 3901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 3961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 4021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 4081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 4141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 4201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 4261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 4321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 4381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 4441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg 4501 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 4561 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 4621 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 4681 gaaaaaccgt ctatcagggc gatggcccac gctggctaag aactcatcag cctccccggt 4741 ccatctaccc acgtaccaat gcaccaattg gccacaatga cggctactac atggtgccat 4801 tccttcctct ttataggaat ggagactacc tcctgtccaa caatgctctt ggatacgagt 4861 acgcctacct gttggaccca ggtcattgca caacaccaga aatgccctct gatctgcaaa 4921 agacgtgaat atctgttcag acacccatat ccactctgtt ccacacaggt cagaggtttg 4981 tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac 5041 aagtacttag ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt 5101 gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga gtattaagga agtaaaagta 5161 aaagcaagaa agaaaactag agattcttgt ttaagctttt aatctcaaaa aacattaaat 5221 gaaatgcata caaggtttta tcctgcttta gaactgtttg tatttaatta tcaaactata 5281 agacagacaa tctaatgcca gtacacgcta ctcaaagttg taaaacctca gatttaactt 5341 cagtagaagc tgattctcaa aattgttagt gtcaagccta gctcttttgg ggctgaaaag 5401 caatcctgca gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag gtgtattagt 5461 cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg tgtctgaaac 5521 acaggccaga tgggccctcg agcaggaaac agctatgacc atgattacgc caagctatca 5581 actttgtata gaaaagttgg cacgcgcaga ttccgtgtgg gcctggtaat atcttaccat 5641 tttaagctat tttaagatta gggaaaaaaa ttaaataaaa ctactttaaa attactttcc 5701 ttaaaaagca acatatttat ttagtttttg tttaactggc tcaagattgt aatattttac 5761 ttttacatgt ttttttaagt acctaaatgt gttcaaaagt aaagtatgtt attgtttttt 5821 ttttttaatg ttagctaagc tttaaattgt aatatattac ttttaaaagt gactttttaa 5881 aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg ttatttctgt gggtttttaa 5941 ttttcatctc aactgggctt acaattttaa cataataaaa atcaaatcaa atcaaatcag 6001 gcccttattt gtcacataca ctcttgtata ctgaaataaa ctaaatgttg agaaaaattt 6061 aaatacacca aaataacgca tacctttttt gtaattacct aagtaaaaca tttagttcct 6121 tttttaagtg actcaagatt gtaatctatt aattgtaaaa gataaaacta gtgatcaatc 6181 tgacgtgctt ctatgcagcc tgtgagatta attgtgatgt ttaattatta aacaatttaa 6241 atggatcaaa catttcacat gtgcttattg tgtccaacaa aaccagcaga agcacttaaa 6301 aatattgcat gacaacatga gtttttaatg taaattatat atacttccta aacgaagtta 6361 atcatatttc taaaataata atgtcattga aatatttaaa taaatgataa gctttagttg 6421 attcaacata aaattgaaag caataaggac tttttttttt tacagcggtg aagcaaaagc 6481 atgtaataat catataatat aataaataaa taaataaata atggctcttg gtgcaaatat 6541 gaacacttta aataaatata gaataaatat gatcaataag tcataacgat gtgttttatg 6601 ttgattataa tttaaacatg tttctaacgc aagttttaaa aactgtttta aagtattatt 6661 ttaagttata agatcaattt gactcaacat tttgtagtga aaatatgttg tgtaaaatta 6721 atataatata cactataaaa atgcagggtt ccacacaata cattaatgtt gtcccaacac 6781 aaatcgatta agttaacaaa tatatgtgga ttgaacataa aaaattaagt tgtcccaatg 6841 aaatctcaag aattgtgttg tttcagctca ttttaaataa gtagttttaa caagtaaaaa 6901 ataacatatt tttgagtgtg tatatatata tattacaaaa tataataata taaaatgtaa 6961 aagatataaa ccatgctaca tatatttaag agctattttg cacattactt ttttgcaagc 7021 cattttagtg ttttttttac aaatatatat tagctaaata tacaatatct taaatataca 7081 attaatcaga catgcaatta atgcatttaa cctacataaa aatacatgat caacatatgt 7141 tttatgctga ttacaactaa atcaaacgta atgaaattaa tttataaaag cgatttaaat 7201 atgtcaatat tttaagttaa aatcacttga aacctactat aaatgaactt aaattaaaaa 7261 aatctccttt aattccaatg caacagtgtt gttttttcag ctgaggaccc ccggtgcaga 7321 tggttggccc ccggcccgct attctaagga agcctttctt aattttaggg cggcgctgca 7381 tgcgctcaga tcagattcac gggtggagag ggttattttc tcttttttat tttttcttgc 7441 ctgctcatat ttgggcctgt gtgaacccca ggctccgcct cggcccccag gagccgctct 7501 tatttaaagg ggatcctcgc cttcagtctt cgtcccagtg gaagcatcag tcttgtgata 7561 acaacttggt gagtcctaca gaccagcgct cgactctagg ctttaaatgc attgatttct 7621 ctggattagc taccgggaga gttttaactt tgcttattag tcttgtgctc gaactgctag 7681 gtttaaagag tgttttctct aatgcatgca tgaagcgaca gctgaaaacc agaaatagag 7741 cgtttgtcct cctgttaaac tctagccttt ctttggcatt tgtcagtccg agtcttcagc 7801 tgccggcagg agattgggtt tcagtaacac ttgctgagca caaactacac gtgctttctt 7861 cagatttaat tcattattaa tcattctaga tttaaatgat gttcttactc attgctgatt 7921 attttagcct tgcggagcag attaaatgag cacgtgtttc attaagattg atttaaatat 7981 aattaatatt tcaccactgt agttttaaaa ttgccttcta agtcatggtt gatttttaat 8041 aggtatcggc tctaattaga ttactatttt ggtttttaaa tgttatttct atttcatttg 8101 taggctcgtt tttaatgtga ttttattaga ttctagtttt aaaatcatct cttagcctta 8161 atgagcagac tgaatgaaca tgtttcatta agattaactt cattataatg aattatttaa 8221 tcattctagt gggttataaa tcagctctat ttaaattaat tgtcgtgttt atttctattt 8281 aatttttgat tattttttca gggtcgtttt tatgtgattt tttcccgttt ctaattttaa 8341 actaattgtt tgatttttat atgattcgac tctttttaaa tgaattgttt tgcttttttt 8401 attgagcata tatatattat acaatttcaa tcaacaattg ttgtgctttt taaatgttgt 8461 ttctatttca tttctagggt cgattttatt cgattctagt tttaaaatcg tgatttttaa 8521 cttctaacct tactgagcag actgaatgaa catgtttcat caagattaac ttcattatag 8581 ttaattattg aatcgtgcta gtggtttata aatcagctct atttaaatta atcatcatca 8641 ttatttctat ttaattttta ttatttcttt aggattattt ttatgtgatt tttttcacgt 8701 tctagtttta aactcaatgc tcattactgc tcacccttgc tgagcagtct ttcgttaaga 8761 ttaaattaat taataaccat tctagtttta aaacagtgtc ttaaggcata gctgatattt 8821 tacaagttta ctatttaata tatctccttt ctttttttag ggtcgatttc atctgatttg 8881 attaattttt attatcttca ctaagctttt tgttctaggt tttattctca gtattattac 8941 tgttatattt gaactgaatt tctcctcata gtttattcat tttatttatt tattttttgt 9001 ttattcgcta ttttgtattt tagtttttaa ttagaaattt tattagaatc gttttgaact 9061 agtttttatg cactctaaca ttaatattaa atcataacta ttatttttat tataatttat 9121 ttgtgtttat ctttttgtgt agctttcatg tgtttttatt ttagttttat tatgtttgca 9181 ttgaattttt tggactagtt tatataacga tatacagaaa agccatgaca acaaatgttt 9241 ttataatacc tattaagtat aagctaatca gatttccact attctgttca aagttaacat 9301 tattagccat tctgatggat catttgacat ttgtcatatg tctttatgaa cactgctctt 9361 tcagatgcca cttttaaagc tgttttttac agtaaatgaa cgtttattat tatgacatta 9421 tttcctaagg cctctagagt acagcaggat gtggcgtgtg cattaccgtg taattatgct 9481 gactgtaaat gtgcatttat agactgtcag catgtctgac tgtgcgctgt tgtgattggt 9541 cctgcacaga ccaag //