LOCUS Exported 9779 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab40c. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab40c SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9779) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9779) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9779 /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(1603..1623) /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(1631..1647) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1663..1797 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1687..1706) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1741..1760 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2469..2485) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2469..2485) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2522..2542) /label=T3 /note="T3 promoter, forward primer" promoter complement(2522..2540) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2561..2577) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2574..2596) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2585..2601 /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(2609..2639) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2654..2675 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2792..2809) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2963..3551) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3043..3062) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3722..4582) /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 4345..4364 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4583..4687) /gene="bla" /label=AmpR promoter primer_bind complement(4850..4871) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5767..5783 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5803..5823 /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 ggtgggcacc cagggaagcc ctgtgaagag 781 ctacgactac ctgctcaagt tcctcctggt cggagacagc gacgtgggca agggagagat 841 cctggatagc cttcaagacg gatctgctga gtctccatat gcctacagca gcggaatcga 901 ctacaagaca accacgatcc tgttggatgg acggagagtt aaactagagc tttgggacac 961 ttcgggacaa ggcaggtttt gcaccatttt caggtcgtac tccaggggag cgcaggggat 1021 cttgttggtg tatgacatca ccaaccgctg gtcatttgat gggatagacc gctggatcag 1081 ggagattgat gagcatgcac ctggagtgcc ccgtattcta gtgggtaacc ggctacacct 1141 ggcgttcaag cgacaggttc ccacagaaca ggcacgggca tacgcagaaa agaatggcat 1201 gacatttttt gaagtgagtc ctctctgcaa cttcaacgtc atcgagtcct ttacagagct 1261 ctcgcgcatc gtcctgatga ggcatgggat ggagaagttc tggaggccca acagagtatt 1321 cagtctgcag gacttgtgct gccgtgccat tgtttcctgc acccctgtgc acctaatcga 1381 taagctgcca cttcctgtgg ccatcaagag ccatttgaag tcattctcca tggcgaacgg 1441 catgaatgct gtcatgatgc acggccgctc gtattcgctg gccaatgccg ccggaggcag 1501 caaaggtaac agtcttaaac gctccaaatc catccggccg ccacaaagtc caccacagaa 1561 ctgcaccagg aacaactgta agatctccta gcctccaggc accaacttta ttatacatag 1621 ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga catgataaga 1681 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt 1741 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac 1801 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa 1861 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga tcagatctgc 1921 gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg ctgctcgcag 1981 ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt ataatttccc 2041 taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac gtcacttcca 2101 aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt tcataggcag 2161 aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt ggtaatagca 2221 agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt aaataaaatt 2281 gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat 2341 tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt aagtacagta 2401 atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta ccttcagcaa 2461 gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt 2521 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt 2581 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 2641 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 2701 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 2761 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 2821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 2881 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 2941 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 3001 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 3061 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 3121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 3181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 3241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 3301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 3361 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 3421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 3481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 3541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 3601 actcacgtta aggcattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 3661 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 3721 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 3781 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 3841 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 3901 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 3961 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 4021 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 4081 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 4141 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 4201 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 4261 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 4321 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 4381 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 4441 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 4501 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 4561 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 4621 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 4681 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa 4741 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 4801 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 4861 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 4921 gggcgatggc ccacgctggc taagaactca tcagcctccc cggtccatct acccacgtac 4981 caatgcacca attggccaca atgacggcta ctacatggtg ccattccttc ctctttatag 5041 gaatggagac tacctcctgt ccaacaatgc tcttggatac gagtacgcct acctgttgga 5101 cccaggtcat tgcacaacac cagaaatgcc ctctgatctg caaaagacgt gaatatctgt 5161 tcagacaccc atatccactc tgttccacac aggtcagagg tttgtccagg agttcttgac 5221 agaggtgtaa aaagtactca aaaattttac tcaagtgaaa gtacaagtac ttagggaaaa 5281 ttttactcaa ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa aagtaaaaaa 5341 gtactccatt aaaattgtac ttgagtatta aggaagtaaa agtaaaagca agaaagaaaa 5401 ctagagattc ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg catacaaggt 5461 tttatcctgc tttagaactg tttgtattta attatcaaac tataagacag acaatctaat 5521 gccagtacac gctactcaaa gttgtaaaac ctcagattta acttcagtag aagctgattc 5581 tcaaaattgt tagtgtcaag cctagctctt ttggggctga aaagcaatcc tgcagtgctg 5641 aaaagcctct cacaggcagc cgatgcggga agaggtgtat tagtcttgat agagaggctg 5701 caaatagcag gaaacgtgag cagagactcc ctggtgtctg aaacacaggc cagatgggcc 5761 ctcgagcagg aaacagctat gaccatgatt acgccaagct atcaactttg tatagaaaag 5821 ttggcacgcg cagattccgt gtgggcctgg taatatctta ccattttaag ctattttaag 5881 attagggaaa aaaattaaat aaaactactt taaaattact ttccttaaaa agcaacatat 5941 ttatttagtt tttgtttaac tggctcaaga ttgtaatatt ttacttttac atgttttttt 6001 aagtacctaa atgtgttcaa aagtaaagta tgttattgtt tttttttttt aatgttagct 6061 aagctttaaa ttgtaatata ttacttttaa aagtgacttt ttaaaaatgt aaataaaaaa 6121 gtaactataa atgtgtaatg catgttattt ctgtgggttt ttaattttca tctcaactgg 6181 gcttacaatt ttaacataat aaaaatcaaa tcaaatcaaa tcaggccctt atttgtcaca 6241 tacactcttg tatactgaaa taaactaaat gttgagaaaa atttaaatac accaaaataa 6301 cgcatacctt ttttgtaatt acctaagtaa aacatttagt tcctttttta agtgactcaa 6361 gattgtaatc tattaattgt aaaagataaa actagtgatc aatctgacgt gcttctatgc 6421 agcctgtgag attaattgtg atgtttaatt attaaacaat ttaaatggat caaacatttc 6481 acatgtgctt attgtgtcca acaaaaccag cagaagcact taaaaatatt gcatgacaac 6541 atgagttttt aatgtaaatt atatatactt cctaaacgaa gttaatcata tttctaaaat 6601 aataatgtca ttgaaatatt taaataaatg ataagcttta gttgattcaa cataaaattg 6661 aaagcaataa ggactttttt tttttacagc ggtgaagcaa aagcatgtaa taatcatata 6721 atataataaa taaataaata aataatggct cttggtgcaa atatgaacac tttaaataaa 6781 tatagaataa atatgatcaa taagtcataa cgatgtgttt tatgttgatt ataatttaaa 6841 catgtttcta acgcaagttt taaaaactgt tttaaagtat tattttaagt tataagatca 6901 atttgactca acattttgta gtgaaaatat gttgtgtaaa attaatataa tatacactat 6961 aaaaatgcag ggttccacac aatacattaa tgttgtccca acacaaatcg attaagttaa 7021 caaatatatg tggattgaac ataaaaaatt aagttgtccc aatgaaatct caagaattgt 7081 gttgtttcag ctcattttaa ataagtagtt ttaacaagta aaaaataaca tatttttgag 7141 tgtgtatata tatatattac aaaatataat aatataaaat gtaaaagata taaaccatgc 7201 tacatatatt taagagctat tttgcacatt acttttttgc aagccatttt agtgtttttt 7261 ttacaaatat atattagcta aatatacaat atcttaaata tacaattaat cagacatgca 7321 attaatgcat ttaacctaca taaaaataca tgatcaacat atgttttatg ctgattacaa 7381 ctaaatcaaa cgtaatgaaa ttaatttata aaagcgattt aaatatgtca atattttaag 7441 ttaaaatcac ttgaaaccta ctataaatga acttaaatta aaaaaatctc ctttaattcc 7501 aatgcaacag tgttgttttt tcagctgagg acccccggtg cagatggttg gcccccggcc 7561 cgctattcta aggaagcctt tcttaatttt agggcggcgc tgcatgcgct cagatcagat 7621 tcacgggtgg agagggttat tttctctttt ttattttttc ttgcctgctc atatttgggc 7681 ctgtgtgaac cccaggctcc gcctcggccc ccaggagccg ctcttattta aaggggatcc 7741 tcgccttcag tcttcgtccc agtggaagca tcagtcttgt gataacaact tggtgagtcc 7801 tacagaccag cgctcgactc taggctttaa atgcattgat ttctctggat tagctaccgg 7861 gagagtttta actttgctta ttagtcttgt gctcgaactg ctaggtttaa agagtgtttt 7921 ctctaatgca tgcatgaagc gacagctgaa aaccagaaat agagcgtttg tcctcctgtt 7981 aaactctagc ctttctttgg catttgtcag tccgagtctt cagctgccgg caggagattg 8041 ggtttcagta acacttgctg agcacaaact acacgtgctt tcttcagatt taattcatta 8101 ttaatcattc tagatttaaa tgatgttctt actcattgct gattatttta gccttgcgga 8161 gcagattaaa tgagcacgtg tttcattaag attgatttaa atataattaa tatttcacca 8221 ctgtagtttt aaaattgcct tctaagtcat ggttgatttt taataggtat cggctctaat 8281 tagattacta ttttggtttt taaatgttat ttctatttca tttgtaggct cgtttttaat 8341 gtgattttat tagattctag ttttaaaatc atctcttagc cttaatgagc agactgaatg 8401 aacatgtttc attaagatta acttcattat aatgaattat ttaatcattc tagtgggtta 8461 taaatcagct ctatttaaat taattgtcgt gtttatttct atttaatttt tgattatttt 8521 ttcagggtcg tttttatgtg attttttccc gtttctaatt ttaaactaat tgtttgattt 8581 ttatatgatt cgactctttt taaatgaatt gttttgcttt ttttattgag catatatata 8641 ttatacaatt tcaatcaaca attgttgtgc tttttaaatg ttgtttctat ttcatttcta 8701 gggtcgattt tattcgattc tagttttaaa atcgtgattt ttaacttcta accttactga 8761 gcagactgaa tgaacatgtt tcatcaagat taacttcatt atagttaatt attgaatcgt 8821 gctagtggtt tataaatcag ctctatttaa attaatcatc atcattattt ctatttaatt 8881 tttattattt ctttaggatt atttttatgt gatttttttc acgttctagt tttaaactca 8941 atgctcatta ctgctcaccc ttgctgagca gtctttcgtt aagattaaat taattaataa 9001 ccattctagt tttaaaacag tgtcttaagg catagctgat attttacaag tttactattt 9061 aatatatctc ctttcttttt ttagggtcga tttcatctga tttgattaat ttttattatc 9121 ttcactaagc tttttgttct aggttttatt ctcagtatta ttactgttat atttgaactg 9181 aatttctcct catagtttat tcattttatt tatttatttt ttgtttattc gctattttgt 9241 attttagttt ttaattagaa attttattag aatcgttttg aactagtttt tatgcactct 9301 aacattaata ttaaatcata actattattt ttattataat ttatttgtgt ttatcttttt 9361 gtgtagcttt catgtgtttt tattttagtt ttattatgtt tgcattgaat tttttggact 9421 agtttatata acgatataca gaaaagccat gacaacaaat gtttttataa tacctattaa 9481 gtataagcta atcagatttc cactattctg ttcaaagtta acattattag ccattctgat 9541 ggatcatttg acatttgtca tatgtcttta tgaacactgc tctttcagat gccactttta 9601 aagctgtttt ttacagtaaa tgaacgttta ttattatgac attatttcct aaggcctcta 9661 gagtacagca ggatgtggcg tgtgcattac cgtgtaatta tgctgactgt aaatgtgcat 9721 ttatagactg tcagcatgtc tgactgtgcg ctgttgtgat tggtcctgca cagaccaag //