LOCUS Exported 9564 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to Rab41. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-Rab41 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9564) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9564) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9564 /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(1388..1408) /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(1416..1432) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1448..1582 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1472..1491) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1526..1545 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2254..2270) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2254..2270) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2307..2327) /label=T3 /note="T3 promoter, forward primer" promoter complement(2307..2325) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2346..2362) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2359..2381) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2370..2386 /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(2394..2424) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2439..2460 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2577..2594) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2748..3336) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2828..2847) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3507..4367) /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 4130..4149 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4368..4472) /gene="bla" /label=AmpR promoter primer_bind complement(4635..4656) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5552..5568 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5588..5608 /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 ggtgtccact acgaccggtg gcggagagtt 781 cggcgaccct ctgcggaaat ttaagctggt tttcctgggc gagcaaagcg tgggcaaaac 841 ttcacttatc accaggttta tgtatgacag ctttgataat acttatcagg caactattgg 901 tatagacttt ctgtcaaaaa ccatgtacct ggaagatcgc acggttcggc tccagctgtg 961 ggacactgca gggcaggagc gcttccgcag cctcattccc agctacatcc gagactccac 1021 tattgctgtg gtggtctatg acatcaccaa tcttaactct ttccagcaaa cctcaaagtg 1081 gatagatgat gtcagaacag agagaggaag cgatgttatt atcatgttgg ttggcaacaa 1141 gacagacctg ggtgataaaa ggcaagtgtc agtagaggca gcagagaaaa aagctcggga 1201 actgggggtg atgtacatag agaccagcgc caaagctggc tataacgtca aacagctgtt 1261 tcgtcgtgtt gctgctgctt tgcctggaat ggatagtaca ccagagaaga gcaaagagga 1321 catgatcgac atcaaactgg agaagcctcc ggagttgccc gttaccgaaa gcagctgctc 1381 gtgctagcaa ctttattata catagttgat aattcactgg ccgtcgtttt acggtaccat 1441 cgatgatgat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat 1501 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat 1561 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca 1621 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga 1681 ttatgatcct ctagatcaga tctgcgaaga tacggccacg ggtgctcttg atcctgtggc 1741 tgattttgga ctgtgctgct cgcagctgct gatgaatcac atacttcctc cattttcttc 1801 cactgattga ctgttataat ttccctaatt tccaggtcaa ggtgctgtgc attgtggtaa 1861 tagatgtgac atgacgtcac ttccaaagga ccaatgaaca tgtctgacca atttcatata 1921 atgtgaaaac gattttcata ggcagaataa ataacattta aattaaactg ggcatcagcg 1981 caattcaatt ggtttggtaa tagcaaggga aaatagaatg aagtgatctc caaaaaataa 2041 gtactttttg actgtaaata aaattgtaag gagtaaaaag tacttttttt tctaaaaaaa 2101 tgtaattaag taaaagtaaa agtattgatt tttaattgta ctcaagtaaa gtaaaaatcc 2161 ccaaaaataa tacttaagta cagtaatcaa gtaaaattac tcaagtactt tacacctctg 2221 gttcttgacc ccctaccttc agcaagccca gcagatccac tagttctaga gcggccgcca 2281 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa 2341 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 2401 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 2461 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 2521 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 2581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 2641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 2701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 2761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 2821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 2881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 2941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 3001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 3061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 3121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 3181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 3241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 3301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 3361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggca ttttggtcat gagattatca 3421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 3481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 3541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 3601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 3661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 3721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 3781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 3841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 3901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 3961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 4021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 4081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 4141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 4201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 4261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 4321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 4381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 4441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa 4501 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 4561 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 4621 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 4681 tcaaagggcg aaaaaccgtc tatcagggcg atggcccacg ctggctaaga actcatcagc 4741 ctccccggtc catctaccca cgtaccaatg caccaattgg ccacaatgac ggctactaca 4801 tggtgccatt ccttcctctt tataggaatg gagactacct cctgtccaac aatgctcttg 4861 gatacgagta cgcctacctg ttggacccag gtcattgcac aacaccagaa atgccctctg 4921 atctgcaaaa gacgtgaata tctgttcaga cacccatatc cactctgttc cacacaggtc 4981 agaggtttgt ccaggagttc ttgacagagg tgtaaaaagt actcaaaaat tttactcaag 5041 tgaaagtaca agtacttagg gaaaatttta ctcaattaaa agtaaaagta tctggctaga 5101 atcttacttg agtaaaagta aaaaagtact ccattaaaat tgtacttgag tattaaggaa 5161 gtaaaagtaa aagcaagaaa gaaaactaga gattcttgtt taagctttta atctcaaaaa 5221 acattaaatg aaatgcatac aaggttttat cctgctttag aactgtttgt atttaattat 5281 caaactataa gacagacaat ctaatgccag tacacgctac tcaaagttgt aaaacctcag 5341 atttaacttc agtagaagct gattctcaaa attgttagtg tcaagcctag ctcttttggg 5401 gctgaaaagc aatcctgcag tgctgaaaag cctctcacag gcagccgatg cgggaagagg 5461 tgtattagtc ttgatagaga ggctgcaaat agcaggaaac gtgagcagag actccctggt 5521 gtctgaaaca caggccagat gggccctcga gcaggaaaca gctatgacca tgattacgcc 5581 aagctatcaa ctttgtatag aaaagttggc acgcgcagat tccgtgtggg cctggtaata 5641 tcttaccatt ttaagctatt ttaagattag ggaaaaaaat taaataaaac tactttaaaa 5701 ttactttcct taaaaagcaa catatttatt tagtttttgt ttaactggct caagattgta 5761 atattttact tttacatgtt tttttaagta cctaaatgtg ttcaaaagta aagtatgtta 5821 ttgttttttt tttttaatgt tagctaagct ttaaattgta atatattact tttaaaagtg 5881 actttttaaa aatgtaaata aaaaagtaac tataaatgtg taatgcatgt tatttctgtg 5941 ggtttttaat tttcatctca actgggctta caattttaac ataataaaaa tcaaatcaaa 6001 tcaaatcagg cccttatttg tcacatacac tcttgtatac tgaaataaac taaatgttga 6061 gaaaaattta aatacaccaa aataacgcat accttttttg taattaccta agtaaaacat 6121 ttagttcctt ttttaagtga ctcaagattg taatctatta attgtaaaag ataaaactag 6181 tgatcaatct gacgtgcttc tatgcagcct gtgagattaa ttgtgatgtt taattattaa 6241 acaatttaaa tggatcaaac atttcacatg tgcttattgt gtccaacaaa accagcagaa 6301 gcacttaaaa atattgcatg acaacatgag tttttaatgt aaattatata tacttcctaa 6361 acgaagttaa tcatatttct aaaataataa tgtcattgaa atatttaaat aaatgataag 6421 ctttagttga ttcaacataa aattgaaagc aataaggact tttttttttt acagcggtga 6481 agcaaaagca tgtaataatc atataatata ataaataaat aaataaataa tggctcttgg 6541 tgcaaatatg aacactttaa ataaatatag aataaatatg atcaataagt cataacgatg 6601 tgttttatgt tgattataat ttaaacatgt ttctaacgca agttttaaaa actgttttaa 6661 agtattattt taagttataa gatcaatttg actcaacatt ttgtagtgaa aatatgttgt 6721 gtaaaattaa tataatatac actataaaaa tgcagggttc cacacaatac attaatgttg 6781 tcccaacaca aatcgattaa gttaacaaat atatgtggat tgaacataaa aaattaagtt 6841 gtcccaatga aatctcaaga attgtgttgt ttcagctcat tttaaataag tagttttaac 6901 aagtaaaaaa taacatattt ttgagtgtgt atatatatat attacaaaat ataataatat 6961 aaaatgtaaa agatataaac catgctacat atatttaaga gctattttgc acattacttt 7021 tttgcaagcc attttagtgt tttttttaca aatatatatt agctaaatat acaatatctt 7081 aaatatacaa ttaatcagac atgcaattaa tgcatttaac ctacataaaa atacatgatc 7141 aacatatgtt ttatgctgat tacaactaaa tcaaacgtaa tgaaattaat ttataaaagc 7201 gatttaaata tgtcaatatt ttaagttaaa atcacttgaa acctactata aatgaactta 7261 aattaaaaaa atctccttta attccaatgc aacagtgttg ttttttcagc tgaggacccc 7321 cggtgcagat ggttggcccc cggcccgcta ttctaaggaa gcctttctta attttagggc 7381 ggcgctgcat gcgctcagat cagattcacg ggtggagagg gttattttct cttttttatt 7441 ttttcttgcc tgctcatatt tgggcctgtg tgaaccccag gctccgcctc ggcccccagg 7501 agccgctctt atttaaaggg gatcctcgcc ttcagtcttc gtcccagtgg aagcatcagt 7561 cttgtgataa caacttggtg agtcctacag accagcgctc gactctaggc tttaaatgca 7621 ttgatttctc tggattagct accgggagag ttttaacttt gcttattagt cttgtgctcg 7681 aactgctagg tttaaagagt gttttctcta atgcatgcat gaagcgacag ctgaaaacca 7741 gaaatagagc gtttgtcctc ctgttaaact ctagcctttc tttggcattt gtcagtccga 7801 gtcttcagct gccggcagga gattgggttt cagtaacact tgctgagcac aaactacacg 7861 tgctttcttc agatttaatt cattattaat cattctagat ttaaatgatg ttcttactca 7921 ttgctgatta ttttagcctt gcggagcaga ttaaatgagc acgtgtttca ttaagattga 7981 tttaaatata attaatattt caccactgta gttttaaaat tgccttctaa gtcatggttg 8041 atttttaata ggtatcggct ctaattagat tactattttg gtttttaaat gttatttcta 8101 tttcatttgt aggctcgttt ttaatgtgat tttattagat tctagtttta aaatcatctc 8161 ttagccttaa tgagcagact gaatgaacat gtttcattaa gattaacttc attataatga 8221 attatttaat cattctagtg ggttataaat cagctctatt taaattaatt gtcgtgttta 8281 tttctattta atttttgatt attttttcag ggtcgttttt atgtgatttt ttcccgtttc 8341 taattttaaa ctaattgttt gatttttata tgattcgact ctttttaaat gaattgtttt 8401 gcttttttta ttgagcatat atatattata caatttcaat caacaattgt tgtgcttttt 8461 aaatgttgtt tctatttcat ttctagggtc gattttattc gattctagtt ttaaaatcgt 8521 gatttttaac ttctaacctt actgagcaga ctgaatgaac atgtttcatc aagattaact 8581 tcattatagt taattattga atcgtgctag tggtttataa atcagctcta tttaaattaa 8641 tcatcatcat tatttctatt taatttttat tatttcttta ggattatttt tatgtgattt 8701 ttttcacgtt ctagttttaa actcaatgct cattactgct cacccttgct gagcagtctt 8761 tcgttaagat taaattaatt aataaccatt ctagttttaa aacagtgtct taaggcatag 8821 ctgatatttt acaagtttac tatttaatat atctcctttc tttttttagg gtcgatttca 8881 tctgatttga ttaattttta ttatcttcac taagcttttt gttctaggtt ttattctcag 8941 tattattact gttatatttg aactgaattt ctcctcatag tttattcatt ttatttattt 9001 attttttgtt tattcgctat tttgtatttt agtttttaat tagaaatttt attagaatcg 9061 ttttgaacta gtttttatgc actctaacat taatattaaa tcataactat tatttttatt 9121 ataatttatt tgtgtttatc tttttgtgta gctttcatgt gtttttattt tagttttatt 9181 atgtttgcat tgaatttttt ggactagttt atataacgat atacagaaaa gccatgacaa 9241 caaatgtttt tataatacct attaagtata agctaatcag atttccacta ttctgttcaa 9301 agttaacatt attagccatt ctgatggatc atttgacatt tgtcatatgt ctttatgaac 9361 actgctcttt cagatgccac ttttaaagct gttttttaca gtaaatgaac gtttattatt 9421 atgacattat ttcctaaggc ctctagagta cagcaggatg tggcgtgtgc attaccgtgt 9481 aattatgctg actgtaaatg tgcatttata gactgtcagc atgtctgact gtgcgctgtt 9541 gtgattggtc ctgcacagac caag //