LOCUS Exported 10230 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to EHD1a-delta-EH. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-EHD1a-dEH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10230) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 10230) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10230 /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(2054..2074) /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(2082..2098) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 2114..2248 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2138..2157) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2192..2211 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2920..2936) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2920..2936) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2973..2993) /label=T3 /note="T3 promoter, forward primer" promoter complement(2973..2991) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3012..3028) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3025..3047) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3036..3052 /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(3060..3090) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3105..3126 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3243..3260) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3414..4002) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3494..3513) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4173..5033) /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 4796..4815 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5034..5138) /gene="bla" /label=AmpR promoter primer_bind complement(5301..5322) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6218..6234 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 6254..6274 /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 ggctttcagc tggtcgaata gagatggtaa 781 gaaagaccca gaacttttcc aaaacgtgtc cgatggactg aagcgccttt accgcaccaa 841 actcttccct ctggaggaca cctatcattt ccacgacttc cattcaccgg cgctcgaaga 901 tgcggatttt gacaacaagc ccatggtgct gctggtggga cagtactcga ccgggaagac 961 caccttcatc aggcatttaa tggagcagga tttccccggg atgaggatag gcccagagcc 1021 gacgacggat tccttcattg cggttatgca cggtgagcaa gaaggcgtca tacctggtaa 1081 tgcgctggtg gtggatccga aaaagccttt ccgaaaactc aacgcatttg gcaacgcttt 1141 cctcaacagg tttatgtgtg cacagttaaa caacccggta cttgagagca tcagtatcat 1201 cgatactcca ggcatcctgt caggggagaa acagaggatc agcagaggct atgactttgc 1261 tgctgttctg gagtggtttg cggagcgcgt ggaccgaatc atcctgctct tcgacgccca 1321 caagctggac atttcggatg agttttcaga ggtgatcaag gccctgaaga accacgagga 1381 caagatgcgc gtggtgctga ataaagcaga tcagatcagc acgcagcagc tcatgagggt 1441 ttatggagcc ctcatgtggt cattgggcaa gatcataaac acccctgagg tggtgcgggt 1501 ttatatcggc tccttctggg ctcagccgct gctcatcgct gaaaaccgga agctgtttga 1561 agcggaggag caggatcttt tccaggatat tcagagtctg ccgagaaacg cagctctgag 1621 gaaactcaac gatctgatca aaagagcacg tcttgctaag gtacatgctt acattatcag 1681 tgctctgaag aaagagatgc cgagcgtgtt tggcaaagac tcaaagaaga aagagcttat 1741 taacaacttg ggcgctatat atgaaaagat ccagaaagag cacaacattt caccaggaga 1801 ctttccaaac cttaagaaaa tgcaggagtt gctatctgtt caggacttca ccaaattccc 1861 tgccatgaag cccaaacttc tagagtcggt ggaagacatg ctaggtaacg acatcgctaa 1921 gctaatggca ctggtgcgta aagaggaggc caccatgccc agccaatcag tgaagggcgg 1981 tgcttttgag ggaatctcaa acggcccgtt cggtcatggg tacggagaag gcgccggcga 2041 aggcatcgac tgacaacttt attatacata gttgataatt cactggccgt cgttttacgg 2101 taccatcgat gatgatccag acatgataag atacattgat gagtttggac aaaccacaac 2161 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt 2221 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca 2281 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat 2341 ggctgattat gatcctctag atcagatctg cgaagatacg gccacgggtg ctcttgatcc 2401 tgtggctgat tttggactgt gctgctcgca gctgctgatg aatcacatac ttcctccatt 2461 ttcttccact gattgactgt tataatttcc ctaatttcca ggtcaaggtg ctgtgcattg 2521 tggtaataga tgtgacatga cgtcacttcc aaaggaccaa tgaacatgtc tgaccaattt 2581 catataatgt gaaaacgatt ttcataggca gaataaataa catttaaatt aaactgggca 2641 tcagcgcaat tcaattggtt tggtaatagc aagggaaaat agaatgaagt gatctccaaa 2701 aaataagtac tttttgactg taaataaaat tgtaaggagt aaaaagtact tttttttcta 2761 aaaaaatgta attaagtaaa agtaaaagta ttgattttta attgtactca agtaaagtaa 2821 aaatccccaa aaataatact taagtacagt aatcaagtaa aattactcaa gtactttaca 2881 cctctggttc ttgaccccct accttcagca agcccagcag atccactagt tctagagcgg 2941 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg 3001 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 3061 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 3121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 3181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 3241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 3301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 3361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 3421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 3481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 3541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 3601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 3661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 3721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 3781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 3841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 3901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 3961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 4021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aaggcatttt ggtcatgaga 4081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 4141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 4201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 4261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 4321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 4381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 4441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 4501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 4561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 4621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 4681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 4741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 4801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 4861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 4921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 4981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 5041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 5101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 5161 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 5221 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg 5281 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact 5341 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccacgctgg ctaagaactc 5401 atcagcctcc ccggtccatc tacccacgta ccaatgcacc aattggccac aatgacggct 5461 actacatggt gccattcctt cctctttata ggaatggaga ctacctcctg tccaacaatg 5521 ctcttggata cgagtacgcc tacctgttgg acccaggtca ttgcacaaca ccagaaatgc 5581 cctctgatct gcaaaagacg tgaatatctg ttcagacacc catatccact ctgttccaca 5641 caggtcagag gtttgtccag gagttcttga cagaggtgta aaaagtactc aaaaatttta 5701 ctcaagtgaa agtacaagta cttagggaaa attttactca attaaaagta aaagtatctg 5761 gctagaatct tacttgagta aaagtaaaaa agtactccat taaaattgta cttgagtatt 5821 aaggaagtaa aagtaaaagc aagaaagaaa actagagatt cttgtttaag cttttaatct 5881 caaaaaacat taaatgaaat gcatacaagg ttttatcctg ctttagaact gtttgtattt 5941 aattatcaaa ctataagaca gacaatctaa tgccagtaca cgctactcaa agttgtaaaa 6001 cctcagattt aacttcagta gaagctgatt ctcaaaattg ttagtgtcaa gcctagctct 6061 tttggggctg aaaagcaatc ctgcagtgct gaaaagcctc tcacaggcag ccgatgcggg 6121 aagaggtgta ttagtcttga tagagaggct gcaaatagca ggaaacgtga gcagagactc 6181 cctggtgtct gaaacacagg ccagatgggc cctcgagcag gaaacagcta tgaccatgat 6241 tacgccaagc tatcaacttt gtatagaaaa gttggcacgc gcagattccg tgtgggcctg 6301 gtaatatctt accattttaa gctattttaa gattagggaa aaaaattaaa taaaactact 6361 ttaaaattac tttccttaaa aagcaacata tttatttagt ttttgtttaa ctggctcaag 6421 attgtaatat tttactttta catgtttttt taagtaccta aatgtgttca aaagtaaagt 6481 atgttattgt tttttttttt taatgttagc taagctttaa attgtaatat attactttta 6541 aaagtgactt tttaaaaatg taaataaaaa agtaactata aatgtgtaat gcatgttatt 6601 tctgtgggtt tttaattttc atctcaactg ggcttacaat tttaacataa taaaaatcaa 6661 atcaaatcaa atcaggccct tatttgtcac atacactctt gtatactgaa ataaactaaa 6721 tgttgagaaa aatttaaata caccaaaata acgcatacct tttttgtaat tacctaagta 6781 aaacatttag ttcctttttt aagtgactca agattgtaat ctattaattg taaaagataa 6841 aactagtgat caatctgacg tgcttctatg cagcctgtga gattaattgt gatgtttaat 6901 tattaaacaa tttaaatgga tcaaacattt cacatgtgct tattgtgtcc aacaaaacca 6961 gcagaagcac ttaaaaatat tgcatgacaa catgagtttt taatgtaaat tatatatact 7021 tcctaaacga agttaatcat atttctaaaa taataatgtc attgaaatat ttaaataaat 7081 gataagcttt agttgattca acataaaatt gaaagcaata aggacttttt ttttttacag 7141 cggtgaagca aaagcatgta ataatcatat aatataataa ataaataaat aaataatggc 7201 tcttggtgca aatatgaaca ctttaaataa atatagaata aatatgatca ataagtcata 7261 acgatgtgtt ttatgttgat tataatttaa acatgtttct aacgcaagtt ttaaaaactg 7321 ttttaaagta ttattttaag ttataagatc aatttgactc aacattttgt agtgaaaata 7381 tgttgtgtaa aattaatata atatacacta taaaaatgca gggttccaca caatacatta 7441 atgttgtccc aacacaaatc gattaagtta acaaatatat gtggattgaa cataaaaaat 7501 taagttgtcc caatgaaatc tcaagaattg tgttgtttca gctcatttta aataagtagt 7561 tttaacaagt aaaaaataac atatttttga gtgtgtatat atatatatta caaaatataa 7621 taatataaaa tgtaaaagat ataaaccatg ctacatatat ttaagagcta ttttgcacat 7681 tacttttttg caagccattt tagtgttttt tttacaaata tatattagct aaatatacaa 7741 tatcttaaat atacaattaa tcagacatgc aattaatgca tttaacctac ataaaaatac 7801 atgatcaaca tatgttttat gctgattaca actaaatcaa acgtaatgaa attaatttat 7861 aaaagcgatt taaatatgtc aatattttaa gttaaaatca cttgaaacct actataaatg 7921 aacttaaatt aaaaaaatct cctttaattc caatgcaaca gtgttgtttt ttcagctgag 7981 gacccccggt gcagatggtt ggcccccggc ccgctattct aaggaagcct ttcttaattt 8041 tagggcggcg ctgcatgcgc tcagatcaga ttcacgggtg gagagggtta ttttctcttt 8101 tttatttttt cttgcctgct catatttggg cctgtgtgaa ccccaggctc cgcctcggcc 8161 cccaggagcc gctcttattt aaaggggatc ctcgccttca gtcttcgtcc cagtggaagc 8221 atcagtcttg tgataacaac ttggtgagtc ctacagacca gcgctcgact ctaggcttta 8281 aatgcattga tttctctgga ttagctaccg ggagagtttt aactttgctt attagtcttg 8341 tgctcgaact gctaggttta aagagtgttt tctctaatgc atgcatgaag cgacagctga 8401 aaaccagaaa tagagcgttt gtcctcctgt taaactctag cctttctttg gcatttgtca 8461 gtccgagtct tcagctgccg gcaggagatt gggtttcagt aacacttgct gagcacaaac 8521 tacacgtgct ttcttcagat ttaattcatt attaatcatt ctagatttaa atgatgttct 8581 tactcattgc tgattatttt agccttgcgg agcagattaa atgagcacgt gtttcattaa 8641 gattgattta aatataatta atatttcacc actgtagttt taaaattgcc ttctaagtca 8701 tggttgattt ttaataggta tcggctctaa ttagattact attttggttt ttaaatgtta 8761 tttctatttc atttgtaggc tcgtttttaa tgtgatttta ttagattcta gttttaaaat 8821 catctcttag ccttaatgag cagactgaat gaacatgttt cattaagatt aacttcatta 8881 taatgaatta tttaatcatt ctagtgggtt ataaatcagc tctatttaaa ttaattgtcg 8941 tgtttatttc tatttaattt ttgattattt tttcagggtc gtttttatgt gattttttcc 9001 cgtttctaat tttaaactaa ttgtttgatt tttatatgat tcgactcttt ttaaatgaat 9061 tgttttgctt tttttattga gcatatatat attatacaat ttcaatcaac aattgttgtg 9121 ctttttaaat gttgtttcta tttcatttct agggtcgatt ttattcgatt ctagttttaa 9181 aatcgtgatt tttaacttct aaccttactg agcagactga atgaacatgt ttcatcaaga 9241 ttaacttcat tatagttaat tattgaatcg tgctagtggt ttataaatca gctctattta 9301 aattaatcat catcattatt tctatttaat ttttattatt tctttaggat tatttttatg 9361 tgattttttt cacgttctag ttttaaactc aatgctcatt actgctcacc cttgctgagc 9421 agtctttcgt taagattaaa ttaattaata accattctag ttttaaaaca gtgtcttaag 9481 gcatagctga tattttacaa gtttactatt taatatatct cctttctttt tttagggtcg 9541 atttcatctg atttgattaa tttttattat cttcactaag ctttttgttc taggttttat 9601 tctcagtatt attactgtta tatttgaact gaatttctcc tcatagttta ttcattttat 9661 ttatttattt tttgtttatt cgctattttg tattttagtt tttaattaga aattttatta 9721 gaatcgtttt gaactagttt ttatgcactc taacattaat attaaatcat aactattatt 9781 tttattataa tttatttgtg tttatctttt tgtgtagctt tcatgtgttt ttattttagt 9841 tttattatgt ttgcattgaa ttttttggac tagtttatat aacgatatac agaaaagcca 9901 tgacaacaaa tgtttttata atacctatta agtataagct aatcagattt ccactattct 9961 gttcaaagtt aacattatta gccattctga tggatcattt gacatttgtc atatgtcttt 10021 atgaacactg ctctttcaga tgccactttt aaagctgttt tttacagtaa atgaacgttt 10081 attattatga cattatttcc taaggcctct agagtacagc aggatgtggc gtgtgcatta 10141 ccgtgtaatt atgctgactg taaatgtgca tttatagact gtcagcatgt ctgactgtgc 10201 gctgttgtga ttggtcctgc acagaccaag //