LOCUS Exported 9576 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab25a. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab25a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9576) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9576) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9576 /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(1400..1420) /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(1428..1444) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1460..1594 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1484..1503) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1538..1557 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2266..2282) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2266..2282) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2319..2339) /label=T3 /note="T3 promoter, forward primer" promoter complement(2319..2337) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2358..2374) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2371..2393) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2382..2398 /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(2406..2436) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2451..2472 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2589..2606) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2760..3348) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2840..2859) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3519..4379) /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 4142..4161 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4380..4484) /gene="bla" /label=AmpR promoter primer_bind complement(4647..4668) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5564..5580 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5600..5620 /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 ggtggggaca gatttagcct acaactttgt 781 cttcaaagtg gttttaattg gagaatcagg ggttggaaag agcaacctgc tgtcaaggtt 841 taccaaaaac gagttcaacc atgacagtcg caccactatt ggcgtggagt tcagcactcg 901 ctctgttcag gtggacagta tcaccatcaa agcccagatc tgggacactg cgggtctaga 961 gcgctacagg gccatcacct cagcgtatta cagaggagca gtgggtgcgc tgctggtcta 1021 tgacatcacc aaacacctga cctatgagag cgtggagcgc tggctgaagg aactgtatga 1081 tcacgctgat cctcatatcg tggtcatgct ggtgggcaat aaaaatgacc tggcaactgt 1141 gcgcactgta cccaccgagg aggccaaaga ctttgcagat gccaatggct tactttttat 1201 ggagacttca gcattagagt cgactaatgt tgaatctgct ttcctcgaag ttctaacagc 1261 aatacaccag aaagttgcta gtaaagaagt gaccagaggc tcgatcagcg ccgtgactct 1321 tggacagaca cacggtccgt ccagggaatt ccaggagagc cgaagctgct gcaaaaactc 1381 ctgaacacac cagaggacac aactttatta tacatagttg ataattcact ggccgtcgtt 1441 ttacggtacc atcgatgatg atccagacat gataagatac attgatgagt ttggacaaac 1501 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 1561 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat 1621 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg 1681 tggtatggct gattatgatc ctctagatca gatctgcgaa gatacggcca cgggtgctct 1741 tgatcctgtg gctgattttg gactgtgctg ctcgcagctg ctgatgaatc acatacttcc 1801 tccattttct tccactgatt gactgttata atttccctaa tttccaggtc aaggtgctgt 1861 gcattgtggt aatagatgtg acatgacgtc acttccaaag gaccaatgaa catgtctgac 1921 caatttcata taatgtgaaa acgattttca taggcagaat aaataacatt taaattaaac 1981 tgggcatcag cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc 2041 tccaaaaaat aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt 2101 tttctaaaaa aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta 2161 aagtaaaaat ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac 2221 tttacacctc tggttcttga ccccctacct tcagcaagcc cagcagatcc actagttcta 2281 gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg 2341 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 2401 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 2461 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 2521 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 2581 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 2641 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 2701 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 2761 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 2821 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 2881 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 2941 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 3001 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 3061 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 3121 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 3181 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 3241 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 3301 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 3361 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg cattttggtc 3421 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 3481 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 3541 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 3601 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 3661 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 3721 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 3781 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 3841 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 3901 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 3961 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 4021 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 4081 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 4141 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 4201 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 4261 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 4321 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 4381 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 4441 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 4501 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa 4561 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat 4621 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg 4681 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca cgctggctaa 4741 gaactcatca gcctccccgg tccatctacc cacgtaccaa tgcaccaatt ggccacaatg 4801 acggctacta catggtgcca ttccttcctc tttataggaa tggagactac ctcctgtcca 4861 acaatgctct tggatacgag tacgcctacc tgttggaccc aggtcattgc acaacaccag 4921 aaatgccctc tgatctgcaa aagacgtgaa tatctgttca gacacccata tccactctgt 4981 tccacacagg tcagaggttt gtccaggagt tcttgacaga ggtgtaaaaa gtactcaaaa 5041 attttactca agtgaaagta caagtactta gggaaaattt tactcaatta aaagtaaaag 5101 tatctggcta gaatcttact tgagtaaaag taaaaaagta ctccattaaa attgtacttg 5161 agtattaagg aagtaaaagt aaaagcaaga aagaaaacta gagattcttg tttaagcttt 5221 taatctcaaa aaacattaaa tgaaatgcat acaaggtttt atcctgcttt agaactgttt 5281 gtatttaatt atcaaactat aagacagaca atctaatgcc agtacacgct actcaaagtt 5341 gtaaaacctc agatttaact tcagtagaag ctgattctca aaattgttag tgtcaagcct 5401 agctcttttg gggctgaaaa gcaatcctgc agtgctgaaa agcctctcac aggcagccga 5461 tgcgggaaga ggtgtattag tcttgataga gaggctgcaa atagcaggaa acgtgagcag 5521 agactccctg gtgtctgaaa cacaggccag atgggccctc gagcaggaaa cagctatgac 5581 catgattacg ccaagctatc aactttgtat agaaaagttg gcacgcgcag attccgtgtg 5641 ggcctggtaa tatcttacca ttttaagcta ttttaagatt agggaaaaaa attaaataaa 5701 actactttaa aattactttc cttaaaaagc aacatattta tttagttttt gtttaactgg 5761 ctcaagattg taatatttta cttttacatg tttttttaag tacctaaatg tgttcaaaag 5821 taaagtatgt tattgttttt tttttttaat gttagctaag ctttaaattg taatatatta 5881 cttttaaaag tgacttttta aaaatgtaaa taaaaaagta actataaatg tgtaatgcat 5941 gttatttctg tgggttttta attttcatct caactgggct tacaatttta acataataaa 6001 aatcaaatca aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa 6061 actaaatgtt gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc 6121 taagtaaaac atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa 6181 agataaaact agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg 6241 tttaattatt aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca 6301 aaaccagcag aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata 6361 tatacttcct aaacgaagtt aatcatattt ctaaaataat aatgtcattg aaatatttaa 6421 ataaatgata agctttagtt gattcaacat aaaattgaaa gcaataagga cttttttttt 6481 ttacagcggt gaagcaaaag catgtaataa tcatataata taataaataa ataaataaat 6541 aatggctctt ggtgcaaata tgaacacttt aaataaatat agaataaata tgatcaataa 6601 gtcataacga tgtgttttat gttgattata atttaaacat gtttctaacg caagttttaa 6661 aaactgtttt aaagtattat tttaagttat aagatcaatt tgactcaaca ttttgtagtg 6721 aaaatatgtt gtgtaaaatt aatataatat acactataaa aatgcagggt tccacacaat 6781 acattaatgt tgtcccaaca caaatcgatt aagttaacaa atatatgtgg attgaacata 6841 aaaaattaag ttgtcccaat gaaatctcaa gaattgtgtt gtttcagctc attttaaata 6901 agtagtttta acaagtaaaa aataacatat ttttgagtgt gtatatatat atattacaaa 6961 atataataat ataaaatgta aaagatataa accatgctac atatatttaa gagctatttt 7021 gcacattact tttttgcaag ccattttagt gtttttttta caaatatata ttagctaaat 7081 atacaatatc ttaaatatac aattaatcag acatgcaatt aatgcattta acctacataa 7141 aaatacatga tcaacatatg ttttatgctg attacaacta aatcaaacgt aatgaaatta 7201 atttataaaa gcgatttaaa tatgtcaata ttttaagtta aaatcacttg aaacctacta 7261 taaatgaact taaattaaaa aaatctcctt taattccaat gcaacagtgt tgttttttca 7321 gctgaggacc cccggtgcag atggttggcc cccggcccgc tattctaagg aagcctttct 7381 taattttagg gcggcgctgc atgcgctcag atcagattca cgggtggaga gggttatttt 7441 ctctttttta ttttttcttg cctgctcata tttgggcctg tgtgaacccc aggctccgcc 7501 tcggccccca ggagccgctc ttatttaaag gggatcctcg ccttcagtct tcgtcccagt 7561 ggaagcatca gtcttgtgat aacaacttgg tgagtcctac agaccagcgc tcgactctag 7621 gctttaaatg cattgatttc tctggattag ctaccgggag agttttaact ttgcttatta 7681 gtcttgtgct cgaactgcta ggtttaaaga gtgttttctc taatgcatgc atgaagcgac 7741 agctgaaaac cagaaataga gcgtttgtcc tcctgttaaa ctctagcctt tctttggcat 7801 ttgtcagtcc gagtcttcag ctgccggcag gagattgggt ttcagtaaca cttgctgagc 7861 acaaactaca cgtgctttct tcagatttaa ttcattatta atcattctag atttaaatga 7921 tgttcttact cattgctgat tattttagcc ttgcggagca gattaaatga gcacgtgttt 7981 cattaagatt gatttaaata taattaatat ttcaccactg tagttttaaa attgccttct 8041 aagtcatggt tgatttttaa taggtatcgg ctctaattag attactattt tggtttttaa 8101 atgttatttc tatttcattt gtaggctcgt ttttaatgtg attttattag attctagttt 8161 taaaatcatc tcttagcctt aatgagcaga ctgaatgaac atgtttcatt aagattaact 8221 tcattataat gaattattta atcattctag tgggttataa atcagctcta tttaaattaa 8281 ttgtcgtgtt tatttctatt taatttttga ttattttttc agggtcgttt ttatgtgatt 8341 ttttcccgtt tctaatttta aactaattgt ttgattttta tatgattcga ctctttttaa 8401 atgaattgtt ttgctttttt tattgagcat atatatatta tacaatttca atcaacaatt 8461 gttgtgcttt ttaaatgttg tttctatttc atttctaggg tcgattttat tcgattctag 8521 ttttaaaatc gtgattttta acttctaacc ttactgagca gactgaatga acatgtttca 8581 tcaagattaa cttcattata gttaattatt gaatcgtgct agtggtttat aaatcagctc 8641 tatttaaatt aatcatcatc attatttcta tttaattttt attatttctt taggattatt 8701 tttatgtgat ttttttcacg ttctagtttt aaactcaatg ctcattactg ctcacccttg 8761 ctgagcagtc tttcgttaag attaaattaa ttaataacca ttctagtttt aaaacagtgt 8821 cttaaggcat agctgatatt ttacaagttt actatttaat atatctcctt tcttttttta 8881 gggtcgattt catctgattt gattaatttt tattatcttc actaagcttt ttgttctagg 8941 ttttattctc agtattatta ctgttatatt tgaactgaat ttctcctcat agtttattca 9001 ttttatttat ttattttttg tttattcgct attttgtatt ttagttttta attagaaatt 9061 ttattagaat cgttttgaac tagtttttat gcactctaac attaatatta aatcataact 9121 attattttta ttataattta tttgtgttta tctttttgtg tagctttcat gtgtttttat 9181 tttagtttta ttatgtttgc attgaatttt ttggactagt ttatataacg atatacagaa 9241 aagccatgac aacaaatgtt tttataatac ctattaagta taagctaatc agatttccac 9301 tattctgttc aaagttaaca ttattagcca ttctgatgga tcatttgaca tttgtcatat 9361 gtctttatga acactgctct ttcagatgcc acttttaaag ctgtttttta cagtaaatga 9421 acgtttatta ttatgacatt atttcctaag gcctctagag tacagcagga tgtggcgtgt 9481 gcattaccgt gtaattatgc tgactgtaaa tgtgcattta tagactgtca gcatgtctga 9541 ctgtgcgctg ttgtgattgg tcctgcacag accaag //