LOCUS Exported 9202 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to the 5' signal sequence from calreticulin and 3' KDEL ER retention sequence. ACCESSION . VERSION . KEYWORDS actc1b- mKate2-KDEL-pA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9202) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9202) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9202 /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" CDS 89..781 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC DLPSKLGHR" protein_bind complement(803..827) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" polyA_signal 829..963 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(1026..1046) /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(1054..1070) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1086..1220 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1892..1908) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1892..1908) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(1945..1965) /label=T3 /note="T3 promoter, forward primer" promoter complement(1945..1963) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1984..2000) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1997..2019) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2008..2024 /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(2032..2062) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2077..2098 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2215..2232) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2386..2974) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2466..2485) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3145..4005) /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 3768..3787 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4006..4110) /gene="bla" /label=AmpR promoter primer_bind complement(4273..4294) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5190..5206 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5226..5246 /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 catgctgcta tccgtgccgt tgctgctcgg 61 cctcctcggc ctggccgtcg ccctcgaggt gagcgagctg attaaggaga acatgcacat 121 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga 181 aggcaagccc tacgagggca cccagaccat gagaatcaag gcggtcgagg gcggccctct 241 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa 301 ccacacccag ggcatccccg acttctttaa gcagtccttc cccgagggct tcacatggga 361 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca 421 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc 481 tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagaccctgt accccgctga 541 cggcggcctg gaaggcagag ccgacatggc cctgaagctc gtgggcgggg gccacctgat 601 ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg 661 cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt 721 cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacag 781 aggatccaag gacgagctgt aaacccagct ttcttgtaca aagtggggga tccagacatg 841 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt 901 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa 961 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt 1021 ttttccaact ttattataca tagttgataa ttcactggcc gtcgttttac ggtaccatcg 1081 atgatgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 1141 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 1201 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 1261 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt 1321 atgatcctct agatcagatc tgcgaagata cggccacggg tgctcttgat cctgtggctg 1381 attttggact gtgctgctcg cagctgctga tgaatcacat acttcctcca ttttcttcca 1441 ctgattgact gttataattt ccctaatttc caggtcaagg tgctgtgcat tgtggtaata 1501 gatgtgacat gacgtcactt ccaaaggacc aatgaacatg tctgaccaat ttcatataat 1561 gtgaaaacga ttttcatagg cagaataaat aacatttaaa ttaaactggg catcagcgca 1621 attcaattgg tttggtaata gcaagggaaa atagaatgaa gtgatctcca aaaaataagt 1681 actttttgac tgtaaataaa attgtaagga gtaaaaagta cttttttttc taaaaaaatg 1741 taattaagta aaagtaaaag tattgatttt taattgtact caagtaaagt aaaaatcccc 1801 aaaaataata cttaagtaca gtaatcaagt aaaattactc aagtacttta cacctctggt 1861 tcttgacccc ctaccttcag caagcccagc agatccacta gttctagagc ggccgccacc 1921 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc 1981 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 2041 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 2101 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 2161 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 2221 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 2281 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 2341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 2401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 2461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 2521 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 2581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 2641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 2701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 2761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 2821 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 2881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 2941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 3001 gtctgacgct cagtggaacg aaaactcacg ttaaggcatt ttggtcatga gattatcaaa 3061 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 3121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 3181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 3241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 3301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 3361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 3421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 3481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 3541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 3601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 3661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 3721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 3781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 3841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 3901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 3961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 4021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 4081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt 4141 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 4201 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 4261 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 4321 aaagggcgaa aaaccgtcta tcagggcgat ggcccacgct ggctaagaac tcatcagcct 4381 ccccggtcca tctacccacg taccaatgca ccaattggcc acaatgacgg ctactacatg 4441 gtgccattcc ttcctcttta taggaatgga gactacctcc tgtccaacaa tgctcttgga 4501 tacgagtacg cctacctgtt ggacccaggt cattgcacaa caccagaaat gccctctgat 4561 ctgcaaaaga cgtgaatatc tgttcagaca cccatatcca ctctgttcca cacaggtcag 4621 aggtttgtcc aggagttctt gacagaggtg taaaaagtac tcaaaaattt tactcaagtg 4681 aaagtacaag tacttaggga aaattttact caattaaaag taaaagtatc tggctagaat 4741 cttacttgag taaaagtaaa aaagtactcc attaaaattg tacttgagta ttaaggaagt 4801 aaaagtaaaa gcaagaaaga aaactagaga ttcttgttta agcttttaat ctcaaaaaac 4861 attaaatgaa atgcatacaa ggttttatcc tgctttagaa ctgtttgtat ttaattatca 4921 aactataaga cagacaatct aatgccagta cacgctactc aaagttgtaa aacctcagat 4981 ttaacttcag tagaagctga ttctcaaaat tgttagtgtc aagcctagct cttttggggc 5041 tgaaaagcaa tcctgcagtg ctgaaaagcc tctcacaggc agccgatgcg ggaagaggtg 5101 tattagtctt gatagagagg ctgcaaatag caggaaacgt gagcagagac tccctggtgt 5161 ctgaaacaca ggccagatgg gccctcgagc aggaaacagc tatgaccatg attacgccaa 5221 gctatcaact ttgtatagaa aagttggcac gcgcagattc cgtgtgggcc tggtaatatc 5281 ttaccatttt aagctatttt aagattaggg aaaaaaatta aataaaacta ctttaaaatt 5341 actttcctta aaaagcaaca tatttattta gtttttgttt aactggctca agattgtaat 5401 attttacttt tacatgtttt tttaagtacc taaatgtgtt caaaagtaaa gtatgttatt 5461 gttttttttt tttaatgtta gctaagcttt aaattgtaat atattacttt taaaagtgac 5521 tttttaaaaa tgtaaataaa aaagtaacta taaatgtgta atgcatgtta tttctgtggg 5581 tttttaattt tcatctcaac tgggcttaca attttaacat aataaaaatc aaatcaaatc 5641 aaatcaggcc cttatttgtc acatacactc ttgtatactg aaataaacta aatgttgaga 5701 aaaatttaaa tacaccaaaa taacgcatac cttttttgta attacctaag taaaacattt 5761 agttcctttt ttaagtgact caagattgta atctattaat tgtaaaagat aaaactagtg 5821 atcaatctga cgtgcttcta tgcagcctgt gagattaatt gtgatgttta attattaaac 5881 aatttaaatg gatcaaacat ttcacatgtg cttattgtgt ccaacaaaac cagcagaagc 5941 acttaaaaat attgcatgac aacatgagtt tttaatgtaa attatatata cttcctaaac 6001 gaagttaatc atatttctaa aataataatg tcattgaaat atttaaataa atgataagct 6061 ttagttgatt caacataaaa ttgaaagcaa taaggacttt ttttttttac agcggtgaag 6121 caaaagcatg taataatcat ataatataat aaataaataa ataaataatg gctcttggtg 6181 caaatatgaa cactttaaat aaatatagaa taaatatgat caataagtca taacgatgtg 6241 ttttatgttg attataattt aaacatgttt ctaacgcaag ttttaaaaac tgttttaaag 6301 tattatttta agttataaga tcaatttgac tcaacatttt gtagtgaaaa tatgttgtgt 6361 aaaattaata taatatacac tataaaaatg cagggttcca cacaatacat taatgttgtc 6421 ccaacacaaa tcgattaagt taacaaatat atgtggattg aacataaaaa attaagttgt 6481 cccaatgaaa tctcaagaat tgtgttgttt cagctcattt taaataagta gttttaacaa 6541 gtaaaaaata acatattttt gagtgtgtat atatatatat tacaaaatat aataatataa 6601 aatgtaaaag atataaacca tgctacatat atttaagagc tattttgcac attacttttt 6661 tgcaagccat tttagtgttt tttttacaaa tatatattag ctaaatatac aatatcttaa 6721 atatacaatt aatcagacat gcaattaatg catttaacct acataaaaat acatgatcaa 6781 catatgtttt atgctgatta caactaaatc aaacgtaatg aaattaattt ataaaagcga 6841 tttaaatatg tcaatatttt aagttaaaat cacttgaaac ctactataaa tgaacttaaa 6901 ttaaaaaaat ctcctttaat tccaatgcaa cagtgttgtt ttttcagctg aggacccccg 6961 gtgcagatgg ttggcccccg gcccgctatt ctaaggaagc ctttcttaat tttagggcgg 7021 cgctgcatgc gctcagatca gattcacggg tggagagggt tattttctct tttttatttt 7081 ttcttgcctg ctcatatttg ggcctgtgtg aaccccaggc tccgcctcgg cccccaggag 7141 ccgctcttat ttaaagggga tcctcgcctt cagtcttcgt cccagtggaa gcatcagtct 7201 tgtgataaca acttggtgag tcctacagac cagcgctcga ctctaggctt taaatgcatt 7261 gatttctctg gattagctac cgggagagtt ttaactttgc ttattagtct tgtgctcgaa 7321 ctgctaggtt taaagagtgt tttctctaat gcatgcatga agcgacagct gaaaaccaga 7381 aatagagcgt ttgtcctcct gttaaactct agcctttctt tggcatttgt cagtccgagt 7441 cttcagctgc cggcaggaga ttgggtttca gtaacacttg ctgagcacaa actacacgtg 7501 ctttcttcag atttaattca ttattaatca ttctagattt aaatgatgtt cttactcatt 7561 gctgattatt ttagccttgc ggagcagatt aaatgagcac gtgtttcatt aagattgatt 7621 taaatataat taatatttca ccactgtagt tttaaaattg ccttctaagt catggttgat 7681 ttttaatagg tatcggctct aattagatta ctattttggt ttttaaatgt tatttctatt 7741 tcatttgtag gctcgttttt aatgtgattt tattagattc tagttttaaa atcatctctt 7801 agccttaatg agcagactga atgaacatgt ttcattaaga ttaacttcat tataatgaat 7861 tatttaatca ttctagtggg ttataaatca gctctattta aattaattgt cgtgtttatt 7921 tctatttaat ttttgattat tttttcaggg tcgtttttat gtgatttttt cccgtttcta 7981 attttaaact aattgtttga tttttatatg attcgactct ttttaaatga attgttttgc 8041 tttttttatt gagcatatat atattataca atttcaatca acaattgttg tgctttttaa 8101 atgttgtttc tatttcattt ctagggtcga ttttattcga ttctagtttt aaaatcgtga 8161 tttttaactt ctaaccttac tgagcagact gaatgaacat gtttcatcaa gattaacttc 8221 attatagtta attattgaat cgtgctagtg gtttataaat cagctctatt taaattaatc 8281 atcatcatta tttctattta atttttatta tttctttagg attattttta tgtgattttt 8341 ttcacgttct agttttaaac tcaatgctca ttactgctca cccttgctga gcagtctttc 8401 gttaagatta aattaattaa taaccattct agttttaaaa cagtgtctta aggcatagct 8461 gatattttac aagtttacta tttaatatat ctcctttctt tttttagggt cgatttcatc 8521 tgatttgatt aatttttatt atcttcacta agctttttgt tctaggtttt attctcagta 8581 ttattactgt tatatttgaa ctgaatttct cctcatagtt tattcatttt atttatttat 8641 tttttgttta ttcgctattt tgtattttag tttttaatta gaaattttat tagaatcgtt 8701 ttgaactagt ttttatgcac tctaacatta atattaaatc ataactatta tttttattat 8761 aatttatttg tgtttatctt tttgtgtagc tttcatgtgt ttttatttta gttttattat 8821 gtttgcattg aattttttgg actagtttat ataacgatat acagaaaagc catgacaaca 8881 aatgttttta taatacctat taagtataag ctaatcagat ttccactatt ctgttcaaag 8941 ttaacattat tagccattct gatggatcat ttgacatttg tcatatgtct ttatgaacac 9001 tgctctttca gatgccactt ttaaagctgt tttttacagt aaatgaacgt ttattattat 9061 gacattattt cctaaggcct ctagagtaca gcaggatgtg gcgtgtgcat taccgtgtaa 9121 ttatgctgac tgtaaatgtg catttataga ctgtcagcat gtctgactgt gcgctgttgt 9181 gattggtcct gcacagacca ag //