LOCUS Exported 9549 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab1ba. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab1ba SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9549) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9549) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9549 /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(1373..1393) /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(1401..1417) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1433..1567 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1457..1476) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1511..1530 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2239..2255) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2239..2255) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2292..2312) /label=T3 /note="T3 promoter, forward primer" promoter complement(2292..2310) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2331..2347) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2344..2366) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2355..2371 /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(2379..2409) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2424..2445 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2562..2579) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2733..3321) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2813..2832) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3492..4352) /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 4115..4134 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4353..4457) /gene="bla" /label=AmpR promoter primer_bind complement(4620..4641) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5537..5553 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5573..5593 /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 ggtgaatcct gaatatgact acctgttcaa 781 gttgctcctc atcggtgact ctggagtggg aaagtcgtgt ctgttgttga gatttgctga 841 tgacacatac actgagagct atatcagcac catcggcgtg gacttcaaga tccgcaccat 901 tgaacttgat ggcaaaacca ttaagctaca gatatgggat actgcagggc aagaaagatt 961 tcgaactatc acctccagtt actatcgagg ggctcatgga atcattgtgg tttatgatgt 1021 gacggatcag gaatcgtata acaatgttaa acagtggctt caggaaatag accgctacgc 1081 aagcgagaat gtcaacaagc tgctggtggg gaacaaatgc gacctcacca ccaagaaagt 1141 agtggactac acaacagcca aggaatttgc tgactcttta gcaattccct tccttgagac 1201 gagcgctaag aatgccacaa atgtggagca ggccttcatg actatggctg ctgagatcaa 1261 gaagcgtatg ggccctggag cgacaaccgg aggagacaag cccaacctga agatcgagag 1321 cactcctgtg cggcagtctg gcggaggctg ctgttgaaca gcttgagcgt gccaacttta 1381 ttatacatag ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga 1441 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg 1501 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa 1561 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga 1621 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga 1681 tcagatctgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg 1741 ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt 1801 ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac 1861 gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt 1921 tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt 1981 ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt 2041 aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa 2101 gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt 2161 aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta 2221 ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc 2281 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg 2341 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 2401 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 2461 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 2521 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 2581 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 2641 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 2701 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 2761 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 2821 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 2881 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 2941 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 3001 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 3061 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 3121 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 3181 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 3241 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 3301 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 3361 tggaacgaaa actcacgtta aggcattttg gtcatgagat tatcaaaaag gatcttcacc 3421 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 3481 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 3541 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 3601 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 3661 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 3721 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 3781 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 3841 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 3901 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 3961 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 4021 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 4081 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 4141 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 4201 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 4261 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 4321 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 4381 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 4441 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata 4501 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 4561 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 4621 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 4681 ccgtctatca gggcgatggc ccacgctggc taagaactca tcagcctccc cggtccatct 4741 acccacgtac caatgcacca attggccaca atgacggcta ctacatggtg ccattccttc 4801 ctctttatag gaatggagac tacctcctgt ccaacaatgc tcttggatac gagtacgcct 4861 acctgttgga cccaggtcat tgcacaacac cagaaatgcc ctctgatctg caaaagacgt 4921 gaatatctgt tcagacaccc atatccactc tgttccacac aggtcagagg tttgtccagg 4981 agttcttgac agaggtgtaa aaagtactca aaaattttac tcaagtgaaa gtacaagtac 5041 ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa 5101 aagtaaaaaa gtactccatt aaaattgtac ttgagtatta aggaagtaaa agtaaaagca 5161 agaaagaaaa ctagagattc ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg 5221 catacaaggt tttatcctgc tttagaactg tttgtattta attatcaaac tataagacag 5281 acaatctaat gccagtacac gctactcaaa gttgtaaaac ctcagattta acttcagtag 5341 aagctgattc tcaaaattgt tagtgtcaag cctagctctt ttggggctga aaagcaatcc 5401 tgcagtgctg aaaagcctct cacaggcagc cgatgcggga agaggtgtat tagtcttgat 5461 agagaggctg caaatagcag gaaacgtgag cagagactcc ctggtgtctg aaacacaggc 5521 cagatgggcc ctcgagcagg aaacagctat gaccatgatt acgccaagct atcaactttg 5581 tatagaaaag ttggcacgcg cagattccgt gtgggcctgg taatatctta ccattttaag 5641 ctattttaag attagggaaa aaaattaaat aaaactactt taaaattact ttccttaaaa 5701 agcaacatat ttatttagtt tttgtttaac tggctcaaga ttgtaatatt ttacttttac 5761 atgttttttt aagtacctaa atgtgttcaa aagtaaagta tgttattgtt tttttttttt 5821 aatgttagct aagctttaaa ttgtaatata ttacttttaa aagtgacttt ttaaaaatgt 5881 aaataaaaaa gtaactataa atgtgtaatg catgttattt ctgtgggttt ttaattttca 5941 tctcaactgg gcttacaatt ttaacataat aaaaatcaaa tcaaatcaaa tcaggccctt 6001 atttgtcaca tacactcttg tatactgaaa taaactaaat gttgagaaaa atttaaatac 6061 accaaaataa cgcatacctt ttttgtaatt acctaagtaa aacatttagt tcctttttta 6121 agtgactcaa gattgtaatc tattaattgt aaaagataaa actagtgatc aatctgacgt 6181 gcttctatgc agcctgtgag attaattgtg atgtttaatt attaaacaat ttaaatggat 6241 caaacatttc acatgtgctt attgtgtcca acaaaaccag cagaagcact taaaaatatt 6301 gcatgacaac atgagttttt aatgtaaatt atatatactt cctaaacgaa gttaatcata 6361 tttctaaaat aataatgtca ttgaaatatt taaataaatg ataagcttta gttgattcaa 6421 cataaaattg aaagcaataa ggactttttt tttttacagc ggtgaagcaa aagcatgtaa 6481 taatcatata atataataaa taaataaata aataatggct cttggtgcaa atatgaacac 6541 tttaaataaa tatagaataa atatgatcaa taagtcataa cgatgtgttt tatgttgatt 6601 ataatttaaa catgtttcta acgcaagttt taaaaactgt tttaaagtat tattttaagt 6661 tataagatca atttgactca acattttgta gtgaaaatat gttgtgtaaa attaatataa 6721 tatacactat aaaaatgcag ggttccacac aatacattaa tgttgtccca acacaaatcg 6781 attaagttaa caaatatatg tggattgaac ataaaaaatt aagttgtccc aatgaaatct 6841 caagaattgt gttgtttcag ctcattttaa ataagtagtt ttaacaagta aaaaataaca 6901 tatttttgag tgtgtatata tatatattac aaaatataat aatataaaat gtaaaagata 6961 taaaccatgc tacatatatt taagagctat tttgcacatt acttttttgc aagccatttt 7021 agtgtttttt ttacaaatat atattagcta aatatacaat atcttaaata tacaattaat 7081 cagacatgca attaatgcat ttaacctaca taaaaataca tgatcaacat atgttttatg 7141 ctgattacaa ctaaatcaaa cgtaatgaaa ttaatttata aaagcgattt aaatatgtca 7201 atattttaag ttaaaatcac ttgaaaccta ctataaatga acttaaatta aaaaaatctc 7261 ctttaattcc aatgcaacag tgttgttttt tcagctgagg acccccggtg cagatggttg 7321 gcccccggcc cgctattcta aggaagcctt tcttaatttt agggcggcgc tgcatgcgct 7381 cagatcagat tcacgggtgg agagggttat tttctctttt ttattttttc ttgcctgctc 7441 atatttgggc ctgtgtgaac cccaggctcc gcctcggccc ccaggagccg ctcttattta 7501 aaggggatcc tcgccttcag tcttcgtccc agtggaagca tcagtcttgt gataacaact 7561 tggtgagtcc tacagaccag cgctcgactc taggctttaa atgcattgat ttctctggat 7621 tagctaccgg gagagtttta actttgctta ttagtcttgt gctcgaactg ctaggtttaa 7681 agagtgtttt ctctaatgca tgcatgaagc gacagctgaa aaccagaaat agagcgtttg 7741 tcctcctgtt aaactctagc ctttctttgg catttgtcag tccgagtctt cagctgccgg 7801 caggagattg ggtttcagta acacttgctg agcacaaact acacgtgctt tcttcagatt 7861 taattcatta ttaatcattc tagatttaaa tgatgttctt actcattgct gattatttta 7921 gccttgcgga gcagattaaa tgagcacgtg tttcattaag attgatttaa atataattaa 7981 tatttcacca ctgtagtttt aaaattgcct tctaagtcat ggttgatttt taataggtat 8041 cggctctaat tagattacta ttttggtttt taaatgttat ttctatttca tttgtaggct 8101 cgtttttaat gtgattttat tagattctag ttttaaaatc atctcttagc cttaatgagc 8161 agactgaatg aacatgtttc attaagatta acttcattat aatgaattat ttaatcattc 8221 tagtgggtta taaatcagct ctatttaaat taattgtcgt gtttatttct atttaatttt 8281 tgattatttt ttcagggtcg tttttatgtg attttttccc gtttctaatt ttaaactaat 8341 tgtttgattt ttatatgatt cgactctttt taaatgaatt gttttgcttt ttttattgag 8401 catatatata ttatacaatt tcaatcaaca attgttgtgc tttttaaatg ttgtttctat 8461 ttcatttcta gggtcgattt tattcgattc tagttttaaa atcgtgattt ttaacttcta 8521 accttactga gcagactgaa tgaacatgtt tcatcaagat taacttcatt atagttaatt 8581 attgaatcgt gctagtggtt tataaatcag ctctatttaa attaatcatc atcattattt 8641 ctatttaatt tttattattt ctttaggatt atttttatgt gatttttttc acgttctagt 8701 tttaaactca atgctcatta ctgctcaccc ttgctgagca gtctttcgtt aagattaaat 8761 taattaataa ccattctagt tttaaaacag tgtcttaagg catagctgat attttacaag 8821 tttactattt aatatatctc ctttcttttt ttagggtcga tttcatctga tttgattaat 8881 ttttattatc ttcactaagc tttttgttct aggttttatt ctcagtatta ttactgttat 8941 atttgaactg aatttctcct catagtttat tcattttatt tatttatttt ttgtttattc 9001 gctattttgt attttagttt ttaattagaa attttattag aatcgttttg aactagtttt 9061 tatgcactct aacattaata ttaaatcata actattattt ttattataat ttatttgtgt 9121 ttatcttttt gtgtagcttt catgtgtttt tattttagtt ttattatgtt tgcattgaat 9181 tttttggact agtttatata acgatataca gaaaagccat gacaacaaat gtttttataa 9241 tacctattaa gtataagcta atcagatttc cactattctg ttcaaagtta acattattag 9301 ccattctgat ggatcatttg acatttgtca tatgtcttta tgaacactgc tctttcagat 9361 gccactttta aagctgtttt ttacagtaaa tgaacgttta ttattatgac attatttcct 9421 aaggcctcta gagtacagca ggatgtggcg tgtgcattac cgtgtaatta tgctgactgt 9481 aaatgtgcat ttatagactg tcagcatgtc tgactgtgcg ctgttgtgat tggtcctgca 9541 cagaccaag //