LOCUS Exported 9552 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab8a. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab8a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9552) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9552) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9552 /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(1376..1396) /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(1404..1420) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1436..1570 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1460..1479) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1514..1533 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2242..2258) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2242..2258) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2295..2315) /label=T3 /note="T3 promoter, forward primer" promoter complement(2295..2313) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2334..2350) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2347..2369) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2358..2374 /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(2382..2412) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2427..2448 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2565..2582) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2736..3324) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2816..2835) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3495..4355) /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 4118..4137 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4356..4460) /gene="bla" /label=AmpR promoter primer_bind complement(4623..4644) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5540..5556 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5576..5596 /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 ggtggcgaag acctacgatt atttgtttaa 781 actgctgtta atcggggatt ccggagtcgg gaagacctgt gtgctgttca gattctcaga 841 ggatgccttt aactcgacat ttatctcgac gataggtatc gatttcaaga tcaggacaat 901 agaactagat ggcaagaaga taaagttaca gatatgggac acggcaggac aggaacgatt 961 tcggacgatc acgacagcat attacagagg tgcaatgggt attatgctgg tttatgatat 1021 caccaatgag aagtcatttg acaacatcaa aaactggatc cgaaacatag aggagcatgc 1081 atcagctgat gtggaaaaga tgattttggg gaacaaatgt gacattaatg agaagaggca 1141 ggtgtctaaa gacagaggcg agaagctggc gttagagtac ggcatcaagt tcatggagac 1201 aagtgcaaag gctaatatca acgttgagaa tgctttctta acactcgcca gagacataaa 1261 agcaaagatg gacacaaaat tggagggaaa caatcctcag ggcagcaacc acggtgtgaa 1321 aattaccact gaacagccga aaaagagcag cttctttcgc tgtgtgctac tgtgacaact 1381 ttattataca tagttgataa ttcactggcc gtcgttttac ggtaccatcg atgatgatcc 1441 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa 1501 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa 1561 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg 1621 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct 1681 agatcagatc tgcgaagata cggccacggg tgctcttgat cctgtggctg attttggact 1741 gtgctgctcg cagctgctga tgaatcacat acttcctcca ttttcttcca ctgattgact 1801 gttataattt ccctaatttc caggtcaagg tgctgtgcat tgtggtaata gatgtgacat 1861 gacgtcactt ccaaaggacc aatgaacatg tctgaccaat ttcatataat gtgaaaacga 1921 ttttcatagg cagaataaat aacatttaaa ttaaactggg catcagcgca attcaattgg 1981 tttggtaata gcaagggaaa atagaatgaa gtgatctcca aaaaataagt actttttgac 2041 tgtaaataaa attgtaagga gtaaaaagta cttttttttc taaaaaaatg taattaagta 2101 aaagtaaaag tattgatttt taattgtact caagtaaagt aaaaatcccc aaaaataata 2161 cttaagtaca gtaatcaagt aaaattactc aagtacttta cacctctggt tcttgacccc 2221 ctaccttcag caagcccagc agatccacta gttctagagc ggccgccacc gcggtggagc 2281 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag 2341 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 2401 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 2461 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 2521 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 2581 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 2641 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 2701 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 2761 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 2821 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 2881 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 2941 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 3001 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 3061 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 3121 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 3181 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 3241 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 3301 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 3361 cagtggaacg aaaactcacg ttaaggcatt ttggtcatga gattatcaaa aaggatcttc 3421 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 3481 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 3541 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 3601 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 3661 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 3721 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 3781 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 3841 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 3901 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 3961 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 4021 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 4081 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 4141 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 4201 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 4261 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 4321 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 4381 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 4441 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta 4501 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg 4561 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg 4621 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa 4681 aaaccgtcta tcagggcgat ggcccacgct ggctaagaac tcatcagcct ccccggtcca 4741 tctacccacg taccaatgca ccaattggcc acaatgacgg ctactacatg gtgccattcc 4801 ttcctcttta taggaatgga gactacctcc tgtccaacaa tgctcttgga tacgagtacg 4861 cctacctgtt ggacccaggt cattgcacaa caccagaaat gccctctgat ctgcaaaaga 4921 cgtgaatatc tgttcagaca cccatatcca ctctgttcca cacaggtcag aggtttgtcc 4981 aggagttctt gacagaggtg taaaaagtac tcaaaaattt tactcaagtg aaagtacaag 5041 tacttaggga aaattttact caattaaaag taaaagtatc tggctagaat cttacttgag 5101 taaaagtaaa aaagtactcc attaaaattg tacttgagta ttaaggaagt aaaagtaaaa 5161 gcaagaaaga aaactagaga ttcttgttta agcttttaat ctcaaaaaac attaaatgaa 5221 atgcatacaa ggttttatcc tgctttagaa ctgtttgtat ttaattatca aactataaga 5281 cagacaatct aatgccagta cacgctactc aaagttgtaa aacctcagat ttaacttcag 5341 tagaagctga ttctcaaaat tgttagtgtc aagcctagct cttttggggc tgaaaagcaa 5401 tcctgcagtg ctgaaaagcc tctcacaggc agccgatgcg ggaagaggtg tattagtctt 5461 gatagagagg ctgcaaatag caggaaacgt gagcagagac tccctggtgt ctgaaacaca 5521 ggccagatgg gccctcgagc aggaaacagc tatgaccatg attacgccaa gctatcaact 5581 ttgtatagaa aagttggcac gcgcagattc cgtgtgggcc tggtaatatc ttaccatttt 5641 aagctatttt aagattaggg aaaaaaatta aataaaacta ctttaaaatt actttcctta 5701 aaaagcaaca tatttattta gtttttgttt aactggctca agattgtaat attttacttt 5761 tacatgtttt tttaagtacc taaatgtgtt caaaagtaaa gtatgttatt gttttttttt 5821 tttaatgtta gctaagcttt aaattgtaat atattacttt taaaagtgac tttttaaaaa 5881 tgtaaataaa aaagtaacta taaatgtgta atgcatgtta tttctgtggg tttttaattt 5941 tcatctcaac tgggcttaca attttaacat aataaaaatc aaatcaaatc aaatcaggcc 6001 cttatttgtc acatacactc ttgtatactg aaataaacta aatgttgaga aaaatttaaa 6061 tacaccaaaa taacgcatac cttttttgta attacctaag taaaacattt agttcctttt 6121 ttaagtgact caagattgta atctattaat tgtaaaagat aaaactagtg atcaatctga 6181 cgtgcttcta tgcagcctgt gagattaatt gtgatgttta attattaaac aatttaaatg 6241 gatcaaacat ttcacatgtg cttattgtgt ccaacaaaac cagcagaagc acttaaaaat 6301 attgcatgac aacatgagtt tttaatgtaa attatatata cttcctaaac gaagttaatc 6361 atatttctaa aataataatg tcattgaaat atttaaataa atgataagct ttagttgatt 6421 caacataaaa ttgaaagcaa taaggacttt ttttttttac agcggtgaag caaaagcatg 6481 taataatcat ataatataat aaataaataa ataaataatg gctcttggtg caaatatgaa 6541 cactttaaat aaatatagaa taaatatgat caataagtca taacgatgtg ttttatgttg 6601 attataattt aaacatgttt ctaacgcaag ttttaaaaac tgttttaaag tattatttta 6661 agttataaga tcaatttgac tcaacatttt gtagtgaaaa tatgttgtgt aaaattaata 6721 taatatacac tataaaaatg cagggttcca cacaatacat taatgttgtc ccaacacaaa 6781 tcgattaagt taacaaatat atgtggattg aacataaaaa attaagttgt cccaatgaaa 6841 tctcaagaat tgtgttgttt cagctcattt taaataagta gttttaacaa gtaaaaaata 6901 acatattttt gagtgtgtat atatatatat tacaaaatat aataatataa aatgtaaaag 6961 atataaacca tgctacatat atttaagagc tattttgcac attacttttt tgcaagccat 7021 tttagtgttt tttttacaaa tatatattag ctaaatatac aatatcttaa atatacaatt 7081 aatcagacat gcaattaatg catttaacct acataaaaat acatgatcaa catatgtttt 7141 atgctgatta caactaaatc aaacgtaatg aaattaattt ataaaagcga tttaaatatg 7201 tcaatatttt aagttaaaat cacttgaaac ctactataaa tgaacttaaa ttaaaaaaat 7261 ctcctttaat tccaatgcaa cagtgttgtt ttttcagctg aggacccccg gtgcagatgg 7321 ttggcccccg gcccgctatt ctaaggaagc ctttcttaat tttagggcgg cgctgcatgc 7381 gctcagatca gattcacggg tggagagggt tattttctct tttttatttt ttcttgcctg 7441 ctcatatttg ggcctgtgtg aaccccaggc tccgcctcgg cccccaggag ccgctcttat 7501 ttaaagggga tcctcgcctt cagtcttcgt cccagtggaa gcatcagtct tgtgataaca 7561 acttggtgag tcctacagac cagcgctcga ctctaggctt taaatgcatt gatttctctg 7621 gattagctac cgggagagtt ttaactttgc ttattagtct tgtgctcgaa ctgctaggtt 7681 taaagagtgt tttctctaat gcatgcatga agcgacagct gaaaaccaga aatagagcgt 7741 ttgtcctcct gttaaactct agcctttctt tggcatttgt cagtccgagt cttcagctgc 7801 cggcaggaga ttgggtttca gtaacacttg ctgagcacaa actacacgtg ctttcttcag 7861 atttaattca ttattaatca ttctagattt aaatgatgtt cttactcatt gctgattatt 7921 ttagccttgc ggagcagatt aaatgagcac gtgtttcatt aagattgatt taaatataat 7981 taatatttca ccactgtagt tttaaaattg ccttctaagt catggttgat ttttaatagg 8041 tatcggctct aattagatta ctattttggt ttttaaatgt tatttctatt tcatttgtag 8101 gctcgttttt aatgtgattt tattagattc tagttttaaa atcatctctt agccttaatg 8161 agcagactga atgaacatgt ttcattaaga ttaacttcat tataatgaat tatttaatca 8221 ttctagtggg ttataaatca gctctattta aattaattgt cgtgtttatt tctatttaat 8281 ttttgattat tttttcaggg tcgtttttat gtgatttttt cccgtttcta attttaaact 8341 aattgtttga tttttatatg attcgactct ttttaaatga attgttttgc tttttttatt 8401 gagcatatat atattataca atttcaatca acaattgttg tgctttttaa atgttgtttc 8461 tatttcattt ctagggtcga ttttattcga ttctagtttt aaaatcgtga tttttaactt 8521 ctaaccttac tgagcagact gaatgaacat gtttcatcaa gattaacttc attatagtta 8581 attattgaat cgtgctagtg gtttataaat cagctctatt taaattaatc atcatcatta 8641 tttctattta atttttatta tttctttagg attattttta tgtgattttt ttcacgttct 8701 agttttaaac tcaatgctca ttactgctca cccttgctga gcagtctttc gttaagatta 8761 aattaattaa taaccattct agttttaaaa cagtgtctta aggcatagct gatattttac 8821 aagtttacta tttaatatat ctcctttctt tttttagggt cgatttcatc tgatttgatt 8881 aatttttatt atcttcacta agctttttgt tctaggtttt attctcagta ttattactgt 8941 tatatttgaa ctgaatttct cctcatagtt tattcatttt atttatttat tttttgttta 9001 ttcgctattt tgtattttag tttttaatta gaaattttat tagaatcgtt ttgaactagt 9061 ttttatgcac tctaacatta atattaaatc ataactatta tttttattat aatttatttg 9121 tgtttatctt tttgtgtagc tttcatgtgt ttttatttta gttttattat gtttgcattg 9181 aattttttgg actagtttat ataacgatat acagaaaagc catgacaaca aatgttttta 9241 taatacctat taagtataag ctaatcagat ttccactatt ctgttcaaag ttaacattat 9301 tagccattct gatggatcat ttgacatttg tcatatgtct ttatgaacac tgctctttca 9361 gatgccactt ttaaagctgt tttttacagt aaatgaacgt ttattattat gacattattt 9421 cctaaggcct ctagagtaca gcaggatgtg gcgtgtgcat taccgtgtaa ttatgctgac 9481 tgtaaatgtg catttataga ctgtcagcat gtctgactgt gcgctgttgt gattggtcct 9541 gcacagacca ag //