LOCUS Exported 9603 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to Rab27b. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-Rab27b SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9603) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9603) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9603 /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(1427..1447) /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(1455..1471) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1487..1621 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1511..1530) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1565..1584 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2293..2309) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2293..2309) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2346..2366) /label=T3 /note="T3 promoter, forward primer" promoter complement(2346..2364) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2385..2401) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2398..2420) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2409..2425 /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(2433..2463) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2478..2499 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2616..2633) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2787..3375) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2867..2886) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3546..4406) /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 4169..4188 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4407..4511) /gene="bla" /label=AmpR promoter primer_bind complement(4674..4695) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5591..5607 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5627..5647 /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 ggtgttcagc tccagcagca ctatgaccga 781 tggagattac gattatctga tcaaactcct ggctttgggg gactccgggg tcggcaagac 841 cactttcctc taccgataca cagataacaa gttcaacccc aaattcatca ccaccgtggg 901 catcgacttc agggaaaaga gagtggttta tacgactaac agccccaatg ggacgacggg 961 gaaaaccttt aaggttcacc tgcagctgtg ggacactgcg ggacaggaga ggttcaggag 1021 tttgaccact gcattcttca gggatgccat gggttttctg ctgatgtttg atctgaccag 1081 ccagcagagc tttctcaacg tcaggaactg gatgagtcag ctccaggcta atgcatattg 1141 tgagaatcca gatatcgtgc tggtgggaaa taaagcagat ctagccgacc agagagaagt 1201 tcaggagaaa caagcgaaag aactcgcaga caaatacggc atcccgtact tcgagacgag 1261 tgcagcgacg ggctctgagg tggacaaggc tgtggtgacg ctgctggatc tggtgatgaa 1321 gcggatggag cagtgtgtgg agaagcccag cgcagacgca aacaccagcg acggcagcag 1381 caaactgagc gccgcacctg cccaacagaa gaagtgtgca tgctgacaac tttattatac 1441 atagttgata attcactggc cgtcgtttta cggtaccatc gatgatgatc cagacatgat 1501 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat 1561 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt 1621 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt 1681 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcctc tagatcagat 1741 ctgcgaagat acggccacgg gtgctcttga tcctgtggct gattttggac tgtgctgctc 1801 gcagctgctg atgaatcaca tacttcctcc attttcttcc actgattgac tgttataatt 1861 tccctaattt ccaggtcaag gtgctgtgca ttgtggtaat agatgtgaca tgacgtcact 1921 tccaaaggac caatgaacat gtctgaccaa tttcatataa tgtgaaaacg attttcatag 1981 gcagaataaa taacatttaa attaaactgg gcatcagcgc aattcaattg gtttggtaat 2041 agcaagggaa aatagaatga agtgatctcc aaaaaataag tactttttga ctgtaaataa 2101 aattgtaagg agtaaaaagt actttttttt ctaaaaaaat gtaattaagt aaaagtaaaa 2161 gtattgattt ttaattgtac tcaagtaaag taaaaatccc caaaaataat acttaagtac 2221 agtaatcaag taaaattact caagtacttt acacctctgg ttcttgaccc cctaccttca 2281 gcaagcccag cagatccact agttctagag cggccgccac cgcggtggag ctccagcttt 2341 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct 2401 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 2461 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 2521 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 2581 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 2641 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 2701 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 2761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 2821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 2881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 2941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 3001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 3061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 3121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 3181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 3241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 3301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 3361 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 3421 gaaaactcac gttaaggcat tttggtcatg agattatcaa aaaggatctt cacctagatc 3481 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 3541 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 3601 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 3661 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 3721 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 3781 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 3841 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 3901 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 3961 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 4021 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 4081 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 4141 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 4201 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 4261 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 4321 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 4381 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 4441 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 4501 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt 4561 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 4621 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 4681 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 4741 atcagggcga tggcccacgc tggctaagaa ctcatcagcc tccccggtcc atctacccac 4801 gtaccaatgc accaattggc cacaatgacg gctactacat ggtgccattc cttcctcttt 4861 ataggaatgg agactacctc ctgtccaaca atgctcttgg atacgagtac gcctacctgt 4921 tggacccagg tcattgcaca acaccagaaa tgccctctga tctgcaaaag acgtgaatat 4981 ctgttcagac acccatatcc actctgttcc acacaggtca gaggtttgtc caggagttct 5041 tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt gaaagtacaa gtacttaggg 5101 aaaattttac tcaattaaaa gtaaaagtat ctggctagaa tcttacttga gtaaaagtaa 5161 aaaagtactc cattaaaatt gtacttgagt attaaggaag taaaagtaaa agcaagaaag 5221 aaaactagag attcttgttt aagcttttaa tctcaaaaaa cattaaatga aatgcataca 5281 aggttttatc ctgctttaga actgtttgta tttaattatc aaactataag acagacaatc 5341 taatgccagt acacgctact caaagttgta aaacctcaga tttaacttca gtagaagctg 5401 attctcaaaa ttgttagtgt caagcctagc tcttttgggg ctgaaaagca atcctgcagt 5461 gctgaaaagc ctctcacagg cagccgatgc gggaagaggt gtattagtct tgatagagag 5521 gctgcaaata gcaggaaacg tgagcagaga ctccctggtg tctgaaacac aggccagatg 5581 ggccctcgag caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga 5641 aaagttggca cgcgcagatt ccgtgtgggc ctggtaatat cttaccattt taagctattt 5701 taagattagg gaaaaaaatt aaataaaact actttaaaat tactttcctt aaaaagcaac 5761 atatttattt agtttttgtt taactggctc aagattgtaa tattttactt ttacatgttt 5821 ttttaagtac ctaaatgtgt tcaaaagtaa agtatgttat tgtttttttt ttttaatgtt 5881 agctaagctt taaattgtaa tatattactt ttaaaagtga ctttttaaaa atgtaaataa 5941 aaaagtaact ataaatgtgt aatgcatgtt atttctgtgg gtttttaatt ttcatctcaa 6001 ctgggcttac aattttaaca taataaaaat caaatcaaat caaatcaggc ccttatttgt 6061 cacatacact cttgtatact gaaataaact aaatgttgag aaaaatttaa atacaccaaa 6121 ataacgcata ccttttttgt aattacctaa gtaaaacatt tagttccttt tttaagtgac 6181 tcaagattgt aatctattaa ttgtaaaaga taaaactagt gatcaatctg acgtgcttct 6241 atgcagcctg tgagattaat tgtgatgttt aattattaaa caatttaaat ggatcaaaca 6301 tttcacatgt gcttattgtg tccaacaaaa ccagcagaag cacttaaaaa tattgcatga 6361 caacatgagt ttttaatgta aattatatat acttcctaaa cgaagttaat catatttcta 6421 aaataataat gtcattgaaa tatttaaata aatgataagc tttagttgat tcaacataaa 6481 attgaaagca ataaggactt ttttttttta cagcggtgaa gcaaaagcat gtaataatca 6541 tataatataa taaataaata aataaataat ggctcttggt gcaaatatga acactttaaa 6601 taaatataga ataaatatga tcaataagtc ataacgatgt gttttatgtt gattataatt 6661 taaacatgtt tctaacgcaa gttttaaaaa ctgttttaaa gtattatttt aagttataag 6721 atcaatttga ctcaacattt tgtagtgaaa atatgttgtg taaaattaat ataatataca 6781 ctataaaaat gcagggttcc acacaataca ttaatgttgt cccaacacaa atcgattaag 6841 ttaacaaata tatgtggatt gaacataaaa aattaagttg tcccaatgaa atctcaagaa 6901 ttgtgttgtt tcagctcatt ttaaataagt agttttaaca agtaaaaaat aacatatttt 6961 tgagtgtgta tatatatata ttacaaaata taataatata aaatgtaaaa gatataaacc 7021 atgctacata tatttaagag ctattttgca cattactttt ttgcaagcca ttttagtgtt 7081 ttttttacaa atatatatta gctaaatata caatatctta aatatacaat taatcagaca 7141 tgcaattaat gcatttaacc tacataaaaa tacatgatca acatatgttt tatgctgatt 7201 acaactaaat caaacgtaat gaaattaatt tataaaagcg atttaaatat gtcaatattt 7261 taagttaaaa tcacttgaaa cctactataa atgaacttaa attaaaaaaa tctcctttaa 7321 ttccaatgca acagtgttgt tttttcagct gaggaccccc ggtgcagatg gttggccccc 7381 ggcccgctat tctaaggaag cctttcttaa ttttagggcg gcgctgcatg cgctcagatc 7441 agattcacgg gtggagaggg ttattttctc ttttttattt tttcttgcct gctcatattt 7501 gggcctgtgt gaaccccagg ctccgcctcg gcccccagga gccgctctta tttaaagggg 7561 atcctcgcct tcagtcttcg tcccagtgga agcatcagtc ttgtgataac aacttggtga 7621 gtcctacaga ccagcgctcg actctaggct ttaaatgcat tgatttctct ggattagcta 7681 ccgggagagt tttaactttg cttattagtc ttgtgctcga actgctaggt ttaaagagtg 7741 ttttctctaa tgcatgcatg aagcgacagc tgaaaaccag aaatagagcg tttgtcctcc 7801 tgttaaactc tagcctttct ttggcatttg tcagtccgag tcttcagctg ccggcaggag 7861 attgggtttc agtaacactt gctgagcaca aactacacgt gctttcttca gatttaattc 7921 attattaatc attctagatt taaatgatgt tcttactcat tgctgattat tttagccttg 7981 cggagcagat taaatgagca cgtgtttcat taagattgat ttaaatataa ttaatatttc 8041 accactgtag ttttaaaatt gccttctaag tcatggttga tttttaatag gtatcggctc 8101 taattagatt actattttgg tttttaaatg ttatttctat ttcatttgta ggctcgtttt 8161 taatgtgatt ttattagatt ctagttttaa aatcatctct tagccttaat gagcagactg 8221 aatgaacatg tttcattaag attaacttca ttataatgaa ttatttaatc attctagtgg 8281 gttataaatc agctctattt aaattaattg tcgtgtttat ttctatttaa tttttgatta 8341 ttttttcagg gtcgttttta tgtgattttt tcccgtttct aattttaaac taattgtttg 8401 atttttatat gattcgactc tttttaaatg aattgttttg ctttttttat tgagcatata 8461 tatattatac aatttcaatc aacaattgtt gtgcttttta aatgttgttt ctatttcatt 8521 tctagggtcg attttattcg attctagttt taaaatcgtg atttttaact tctaacctta 8581 ctgagcagac tgaatgaaca tgtttcatca agattaactt cattatagtt aattattgaa 8641 tcgtgctagt ggtttataaa tcagctctat ttaaattaat catcatcatt atttctattt 8701 aatttttatt atttctttag gattattttt atgtgatttt tttcacgttc tagttttaaa 8761 ctcaatgctc attactgctc acccttgctg agcagtcttt cgttaagatt aaattaatta 8821 ataaccattc tagttttaaa acagtgtctt aaggcatagc tgatatttta caagtttact 8881 atttaatata tctcctttct ttttttaggg tcgatttcat ctgatttgat taatttttat 8941 tatcttcact aagctttttg ttctaggttt tattctcagt attattactg ttatatttga 9001 actgaatttc tcctcatagt ttattcattt tatttattta ttttttgttt attcgctatt 9061 ttgtatttta gtttttaatt agaaatttta ttagaatcgt tttgaactag tttttatgca 9121 ctctaacatt aatattaaat cataactatt atttttatta taatttattt gtgtttatct 9181 ttttgtgtag ctttcatgtg tttttatttt agttttatta tgtttgcatt gaattttttg 9241 gactagttta tataacgata tacagaaaag ccatgacaac aaatgttttt ataataccta 9301 ttaagtataa gctaatcaga tttccactat tctgttcaaa gttaacatta ttagccattc 9361 tgatggatca tttgacattt gtcatatgtc tttatgaaca ctgctctttc agatgccact 9421 tttaaagctg ttttttacag taaatgaacg tttattatta tgacattatt tcctaaggcc 9481 tctagagtac agcaggatgt ggcgtgtgca ttaccgtgta attatgctga ctgtaaatgt 9541 gcatttatag actgtcagca tgtctgactg tgcgctgttg tgattggtcc tgcacagacc 9601 aag //