LOCUS Exported 9600 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab20. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab20 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9600) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9600) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9600 /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(1424..1444) /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(1452..1468) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1484..1618 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1508..1527) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1562..1581 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2290..2306) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2290..2306) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2343..2363) /label=T3 /note="T3 promoter, forward primer" promoter complement(2343..2361) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2382..2398) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2395..2417) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2406..2422 /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(2430..2460) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2475..2496 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2613..2630) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2784..3372) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2864..2883) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3543..4403) /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 4166..4185 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4404..4508) /gene="bla" /label=AmpR promoter primer_bind complement(4671..4692) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5588..5604 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5624..5644 /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 ggtgaacgtt ggaaagactt ctctcctcca 781 cagatacacg gagaggaagt ttaaggacac gataagcacc gtcggagggg cgtttttcct 841 caaacagtgg ggtccctaca acatttccat atgggatact gcaggtcgag agcagtttca 901 cggtctgggc tccatgtact gccgtggcgc agctgctgtc atcctaacat atgatgtcac 961 caactggcag agcctggcgg agctggagaa ccgcttcctg tccttgactg acactgccaa 1021 cgcagactgc atcttcgccc ttgtggggaa caaagcggac ctgaccgacc cgttcgctca 1081 agagcctgac agcgagagtg ggagcgaccc gcagcctgta gcctcctgcc cgactcctcc 1141 tgccagccct gtccagcaca aacaggtgag cagagaggac gctgtggctc tgtacgcccg 1201 cgttctccga tacaaaggcc tggacgagaa gacctgcctg cccgccgaga ggatgtgctt 1261 cgagaccagc gctaaaacag gttacaacgt ggacgctttg ttcgagacct tatttgattt 1321 agtactgcca tccattctga agaagaggag cgagggtgaa gccactgtag atctggagac 1381 gccactagag gtgagaagga ctaaagctgg ctgctgtgca tagcaacttt attatacata 1441 gttgataatt cactggccgt cgttttacgg taccatcgat gatgatccag acatgataag 1501 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg 1561 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa 1621 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta 1681 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctctag atcagatctg 1741 cgaagatacg gccacgggtg ctcttgatcc tgtggctgat tttggactgt gctgctcgca 1801 gctgctgatg aatcacatac ttcctccatt ttcttccact gattgactgt tataatttcc 1861 ctaatttcca ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga cgtcacttcc 1921 aaaggaccaa tgaacatgtc tgaccaattt catataatgt gaaaacgatt ttcataggca 1981 gaataaataa catttaaatt aaactgggca tcagcgcaat tcaattggtt tggtaatagc 2041 aagggaaaat agaatgaagt gatctccaaa aaataagtac tttttgactg taaataaaat 2101 tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa agtaaaagta 2161 ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact taagtacagt 2221 aatcaagtaa aattactcaa gtactttaca cctctggttc ttgaccccct accttcagca 2281 agcccagcag atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt 2341 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg 2401 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 2461 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 2521 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 2581 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 2641 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 2701 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 2761 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 2821 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 2881 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 2941 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 3001 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 3061 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 3121 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 3181 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 3241 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 3301 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 3361 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 3421 aactcacgtt aaggcatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 3481 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 3541 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 3601 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 3661 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 3721 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 3781 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 3841 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 3901 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 3961 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 4021 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 4081 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 4141 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 4201 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 4261 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 4321 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 4381 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 4441 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 4501 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa 4561 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 4621 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 4681 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 4741 agggcgatgg cccacgctgg ctaagaactc atcagcctcc ccggtccatc tacccacgta 4801 ccaatgcacc aattggccac aatgacggct actacatggt gccattcctt cctctttata 4861 ggaatggaga ctacctcctg tccaacaatg ctcttggata cgagtacgcc tacctgttgg 4921 acccaggtca ttgcacaaca ccagaaatgc cctctgatct gcaaaagacg tgaatatctg 4981 ttcagacacc catatccact ctgttccaca caggtcagag gtttgtccag gagttcttga 5041 cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta cttagggaaa 5101 attttactca attaaaagta aaagtatctg gctagaatct tacttgagta aaagtaaaaa 5161 agtactccat taaaattgta cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa 5221 actagagatt cttgtttaag cttttaatct caaaaaacat taaatgaaat gcatacaagg 5281 ttttatcctg ctttagaact gtttgtattt aattatcaaa ctataagaca gacaatctaa 5341 tgccagtaca cgctactcaa agttgtaaaa cctcagattt aacttcagta gaagctgatt 5401 ctcaaaattg ttagtgtcaa gcctagctct tttggggctg aaaagcaatc ctgcagtgct 5461 gaaaagcctc tcacaggcag ccgatgcggg aagaggtgta ttagtcttga tagagaggct 5521 gcaaatagca ggaaacgtga gcagagactc cctggtgtct gaaacacagg ccagatgggc 5581 cctcgagcag gaaacagcta tgaccatgat tacgccaagc tatcaacttt gtatagaaaa 5641 gttggcacgc gcagattccg tgtgggcctg gtaatatctt accattttaa gctattttaa 5701 gattagggaa aaaaattaaa taaaactact ttaaaattac tttccttaaa aagcaacata 5761 tttatttagt ttttgtttaa ctggctcaag attgtaatat tttactttta catgtttttt 5821 taagtaccta aatgtgttca aaagtaaagt atgttattgt tttttttttt taatgttagc 5881 taagctttaa attgtaatat attactttta aaagtgactt tttaaaaatg taaataaaaa 5941 agtaactata aatgtgtaat gcatgttatt tctgtgggtt tttaattttc atctcaactg 6001 ggcttacaat tttaacataa taaaaatcaa atcaaatcaa atcaggccct tatttgtcac 6061 atacactctt gtatactgaa ataaactaaa tgttgagaaa aatttaaata caccaaaata 6121 acgcatacct tttttgtaat tacctaagta aaacatttag ttcctttttt aagtgactca 6181 agattgtaat ctattaattg taaaagataa aactagtgat caatctgacg tgcttctatg 6241 cagcctgtga gattaattgt gatgtttaat tattaaacaa tttaaatgga tcaaacattt 6301 cacatgtgct tattgtgtcc aacaaaacca gcagaagcac ttaaaaatat tgcatgacaa 6361 catgagtttt taatgtaaat tatatatact tcctaaacga agttaatcat atttctaaaa 6421 taataatgtc attgaaatat ttaaataaat gataagcttt agttgattca acataaaatt 6481 gaaagcaata aggacttttt ttttttacag cggtgaagca aaagcatgta ataatcatat 6541 aatataataa ataaataaat aaataatggc tcttggtgca aatatgaaca ctttaaataa 6601 atatagaata aatatgatca ataagtcata acgatgtgtt ttatgttgat tataatttaa 6661 acatgtttct aacgcaagtt ttaaaaactg ttttaaagta ttattttaag ttataagatc 6721 aatttgactc aacattttgt agtgaaaata tgttgtgtaa aattaatata atatacacta 6781 taaaaatgca gggttccaca caatacatta atgttgtccc aacacaaatc gattaagtta 6841 acaaatatat gtggattgaa cataaaaaat taagttgtcc caatgaaatc tcaagaattg 6901 tgttgtttca gctcatttta aataagtagt tttaacaagt aaaaaataac atatttttga 6961 gtgtgtatat atatatatta caaaatataa taatataaaa tgtaaaagat ataaaccatg 7021 ctacatatat ttaagagcta ttttgcacat tacttttttg caagccattt tagtgttttt 7081 tttacaaata tatattagct aaatatacaa tatcttaaat atacaattaa tcagacatgc 7141 aattaatgca tttaacctac ataaaaatac atgatcaaca tatgttttat gctgattaca 7201 actaaatcaa acgtaatgaa attaatttat aaaagcgatt taaatatgtc aatattttaa 7261 gttaaaatca cttgaaacct actataaatg aacttaaatt aaaaaaatct cctttaattc 7321 caatgcaaca gtgttgtttt ttcagctgag gacccccggt gcagatggtt ggcccccggc 7381 ccgctattct aaggaagcct ttcttaattt tagggcggcg ctgcatgcgc tcagatcaga 7441 ttcacgggtg gagagggtta ttttctcttt tttatttttt cttgcctgct catatttggg 7501 cctgtgtgaa ccccaggctc cgcctcggcc cccaggagcc gctcttattt aaaggggatc 7561 ctcgccttca gtcttcgtcc cagtggaagc atcagtcttg tgataacaac ttggtgagtc 7621 ctacagacca gcgctcgact ctaggcttta aatgcattga tttctctgga ttagctaccg 7681 ggagagtttt aactttgctt attagtcttg tgctcgaact gctaggttta aagagtgttt 7741 tctctaatgc atgcatgaag cgacagctga aaaccagaaa tagagcgttt gtcctcctgt 7801 taaactctag cctttctttg gcatttgtca gtccgagtct tcagctgccg gcaggagatt 7861 gggtttcagt aacacttgct gagcacaaac tacacgtgct ttcttcagat ttaattcatt 7921 attaatcatt ctagatttaa atgatgttct tactcattgc tgattatttt agccttgcgg 7981 agcagattaa atgagcacgt gtttcattaa gattgattta aatataatta atatttcacc 8041 actgtagttt taaaattgcc ttctaagtca tggttgattt ttaataggta tcggctctaa 8101 ttagattact attttggttt ttaaatgtta tttctatttc atttgtaggc tcgtttttaa 8161 tgtgatttta ttagattcta gttttaaaat catctcttag ccttaatgag cagactgaat 8221 gaacatgttt cattaagatt aacttcatta taatgaatta tttaatcatt ctagtgggtt 8281 ataaatcagc tctatttaaa ttaattgtcg tgtttatttc tatttaattt ttgattattt 8341 tttcagggtc gtttttatgt gattttttcc cgtttctaat tttaaactaa ttgtttgatt 8401 tttatatgat tcgactcttt ttaaatgaat tgttttgctt tttttattga gcatatatat 8461 attatacaat ttcaatcaac aattgttgtg ctttttaaat gttgtttcta tttcatttct 8521 agggtcgatt ttattcgatt ctagttttaa aatcgtgatt tttaacttct aaccttactg 8581 agcagactga atgaacatgt ttcatcaaga ttaacttcat tatagttaat tattgaatcg 8641 tgctagtggt ttataaatca gctctattta aattaatcat catcattatt tctatttaat 8701 ttttattatt tctttaggat tatttttatg tgattttttt cacgttctag ttttaaactc 8761 aatgctcatt actgctcacc cttgctgagc agtctttcgt taagattaaa ttaattaata 8821 accattctag ttttaaaaca gtgtcttaag gcatagctga tattttacaa gtttactatt 8881 taatatatct cctttctttt tttagggtcg atttcatctg atttgattaa tttttattat 8941 cttcactaag ctttttgttc taggttttat tctcagtatt attactgtta tatttgaact 9001 gaatttctcc tcatagttta ttcattttat ttatttattt tttgtttatt cgctattttg 9061 tattttagtt tttaattaga aattttatta gaatcgtttt gaactagttt ttatgcactc 9121 taacattaat attaaatcat aactattatt tttattataa tttatttgtg tttatctttt 9181 tgtgtagctt tcatgtgttt ttattttagt tttattatgt ttgcattgaa ttttttggac 9241 tagtttatat aacgatatac agaaaagcca tgacaacaaa tgtttttata atacctatta 9301 agtataagct aatcagattt ccactattct gttcaaagtt aacattatta gccattctga 9361 tggatcattt gacatttgtc atatgtcttt atgaacactg ctctttcaga tgccactttt 9421 aaagctgttt tttacagtaa atgaacgttt attattatga cattatttcc taaggcctct 9481 agagtacagc aggatgtggc gtgtgcatta ccgtgtaatt atgctgactg taaatgtgca 9541 tttatagact gtcagcatgt ctgactgtgc gctgttgtga ttggtcctgc acagaccaag //