LOCUS Exported 10331 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to CD44a. ACCESSION . VERSION . KEYWORDS actc1b-CD44a-mKate2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10331) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 10331) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10331 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(144..165) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1061..1077 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 1097..1117 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" protein_bind 5092..5116 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(6300..6324) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 6326..7024 /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" polyA_signal 7146..7227 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(7228..7248) /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(7256..7272) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 7288..7422 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7312..7331) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8094..8110) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8094..8110) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(8147..8167) /label=T3 /note="T3 promoter, forward primer" promoter complement(8147..8165) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(8186..8202) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8199..8221) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8210..8226 /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(8234..8264) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8279..8300 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8417..8434) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8588..9176) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8668..8687) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9347..10207) /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 9970..9989 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10208..10312) /gene="bla" /label=AmpR promoter ORIGIN 1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa 241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg 301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca 361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa 421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc 481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt 541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat 601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt 661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa 721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta 781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta 841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc 901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc 961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga 1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat 1081 gattacgcca agctatcaac tttgtataga aaagttggca cgcgcagatt ccgtgtgggc 1141 ctggtaatat cttaccattt taagctattt taagattagg gaaaaaaatt aaataaaact 1201 actttaaaat tactttcctt aaaaagcaac atatttattt agtttttgtt taactggctc 1261 aagattgtaa tattttactt ttacatgttt tttttaagta cctaaatgtg ttcaaaagta 1321 aagtatgtta ttgttttttt ttttttaatg ttagctaagc tttaaattgt aatatattac 1381 ttttaaaagt gactttttaa aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg 1441 ttatttctgt gggtttttta attttcatct caactgggct tacaatttta acataataaa 1501 aatcaaatca aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa 1561 actaaatgtt gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc 1621 taagtaaaac atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa 1681 agataaaact agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg 1741 tttaattatt aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca 1801 aaaccagcag aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata 1861 tatacttcct aaacgaagtt aatcatattt tctaaaataa taatgtcatt gaaatattta 1921 aataaatgat aanctttagt tgattcaaca taaaattgaa agcaataagg actttttttt 1981 ttttacagcg gtgaagcaaa agcntgnaaa taatcatata aatataataa ataaataaat 2041 aaaataatgg ttcttggtgc aaatatgaac actttaaata aatatagaat aaatatgatc 2101 aataagtcat aacgacgtgt tttatgttga ttataattta aacatgtttc taacgcaagt 2161 tttaaaaact gttttaaagt attattttaa gttataagat caatttgact caacattttg 2221 tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc aaggttccac 2281 acaattcatt catgttgtcc caacacaaat cgattaagtt aacaaatata tgtggattga 2341 acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc agctcatttt 2401 aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata tatatatatt 2461 tatatattac aaaatataat aatataaaat gtaaaagata taaaccatgc tacatatatt 2521 taagagctat tttgcacatt acttttttgc aagccatttg agtgtttttt tacaaatata 2581 tattagctaa atatacaata tcttaaatat acaattaatc agacatgcaa aaaatgcatt 2641 taacctacat aaaaatacat gatcaacata tgttttatgc tgattacaac taaatcaaac 2701 gtaatgaaat taatttataa aagcgattta aatatgtcaa tattttaagt taaaatcact 2761 tgaaacctac tataaatgaa cttaaattta aaaaatctcc tttaattcca atgcagcact 2821 gttgtttttt cagctgagga cccccggtgc agatggttgg cccccggccc gctattctaa 2881 ggaagccttt cttaatttta gggcggcgct gcatgcgctc agatcagatt cacgggtgga 2941 gagggttatt ttctcttttt tattttttct tgcctgctca tatttgggcc tgtgtgaacc 3001 ccaggctccg cctcggcccc caggagccgc tcttatttaa aggggatcct cgccttcagt 3061 cttcgtccca gtggaagcat cagtcttgtg ataacaactt ggtgagtcct acagaccagc 3121 gctcgactct aggctttaaa tgcattgatt tctctggatt agctaccggg agagttttaa 3181 ctttgcttat tagtcttgtg cttgaactgc taggtttaaa gagggttttc tctaatgcat 3241 gcatgaagcg acagctgaaa accagaaata gagcgtttgt cctcctgtta aactctagcc 3301 tttctttggc atttgtcagt ccgagtcttc agctgccggc aggagattgg gtttcagtaa 3361 cacttgctga gcacaaacta cacgtgcttc cttcagattt aattcattat taatcattct 3421 agatttaaat gatcttctta ctcattgctg attattttag ccttgcggag cagattaaat 3481 gagcacgtgt ttattaagat tgatttaaat ataattaata tttcaccact gtagttttaa 3541 aattgccttc taagtcatgg ttgattttta ataggtatcg gctctaatta gattactatt 3601 ttggttttta aatgttattt ctatttcatt tgtaggctcg tttttaatgt gattttatta 3661 gattctagtt ttaaaatcat cttttagcct tactgagcag actgaatgaa catgtttcat 3721 taagattaac ttcattatag ttaattattt aatcattcta gtggattata aatcagctct 3781 atttaaattg attgtcgtgt ttatttctat ttgatttttg attatttttt aaggtcgttt 3841 ttatgtgatt tttccccgat tctaatttta aactaattgt ttgattttta tatgattcga 3901 ctctttttaa atgaattgtt ttgctttttt tattgagcat atatatatat tatacaattt 3961 caatcaacaa ttgttgtgct ttttaaatgt tgtttctatt tcatttctag ggtcgatttt 4021 attcgattct agttttaaaa tcgtgatttc taactcctaa ccttactgag cagactgaat 4081 gaacatgttt catcaagatt aacttcatta tagttaatta ttgaatcggg ctagtggttt 4141 ataaatcagc tctattataa attaattgtc atcattattt ctatttaatt tttattattt 4201 ttttaggatt atttttaagg gatttttttc acgttctagt tttaaactca atgctcatta 4261 ctgctcaccc ttgctgagca gtctttcgtt aagattaaat taattaataa cccattctag 4321 ttttaaaaca gtgtcttaag gcatagctga tattttacaa gtttactatt taatatatct 4381 ccatttcttt ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa 4441 gctttttgtt ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc 4501 ctcatagttt attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt 4561 ttttaattag aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa 4621 tattaaatca taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct 4681 ttcatgtgtt tttattttag ttttattatg tttgcattga attttttgga ctagtttata 4741 taacgatata cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc 4801 taatcagatt tccactattc tgttcaaagt taacattatt agccattctg atggatcatt 4861 tgacatttgt catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt 4921 ttttacagta aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag 4981 caggatgtgg cgtgtgcatt actgtgtaat tatgctgact gtaaatgtgc atttatagac 5041 tgtcagcatg tctgactgtg cgctgttgtg atttgtcctg cacagaccaa ggcaagtttg 5101 tacaaaaaag caggctgcca ccatgtggac tttattattt gtagtgtttg ctactggatc 5161 accagctgta cttcaggcag ccccggttca gggtaacagt gggagatgca gttttgctgg 5221 tgttttccac atcgagggaa tcagtcgtta ctctcttacc tttcagaagg ctggcgaact 5281 gtgtcagagt ttaggttaca gacttgcaac acaagaacag gtcactgaag cctataaaaa 5341 aggcttgaga acttgcagat acggctggat tgatggtcaa aatgtcacgt tcctcccaca 5401 cattaatggt ccaaactgtg actccagctc tgctgagatc accttccatg ctaaagcagg 5461 agaatttctt tctgatgtct actgttttga cccatcagat tcatcaccaa actgtgatga 5521 tgttaaatct gaggatgtta tgaatggaaa cgcagccaga gacagttcat tctatgcaaa 5581 accacatgaa gacttcatta cagaattcaa gacagaaggg tttttggtag atgtggaaga 5641 aatattggaa agagtaaaaa gagaaacatc tcttctgagt gacatgaaac cctcaacagt 5701 gccctctatg aaaaaaattg aaggcatcaa ggaccctaca agcaagcctc atattggaaa 5761 gacccccagt gttttgctat ttgatacaga gggcagtgga tcgggattgg ttgatcctga 5821 gcctagccat ttcacaacaa gtccggttac tacagagaca gcgaagagtg atttaattga 5881 aaacgttatt gaagagaatg aagagatggt ggttccagta gagacttcaa atgtgaaccc 5941 cattcaagtc aaggaaccaa gacgagtgaa tgtattttca acaacagaaa gtaatgcgaa 6001 ggagatggtt tcagatggat cgtcttctac ttggatgata attactgcat tctgtgtgtt 6061 tattggagtc atagtatgca tatttgtggc tattggcact agagacaagt ggtacggacc 6121 gagtaagagt gcagacatca caactgaaaa gaacaacaaa gactacagta aaacagaaac 6181 gctgccgctg tcagagaaag agcaggagat tgtggcactg atgaatgtta cgaacatgaa 6241 aaatgggcaa acagtggaca taacagctgc agatgaacac gaaaaagaat atttaatgca 6301 cccagctttc ttgtacaaag tggctatggt gagcgagctg attaaggaga acatgcacat 6361 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga 6421 aggcaagccc tacgagggca cccagaccat gagaatcaag gcggtcgagg gcggccctct 6481 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa 6541 ccacacccag ggcatccccg acttctttaa gcagtccttc cccgagggct tcacatggga 6601 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca 6661 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc 6721 tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagaccctgt accccgctga 6781 cggcggcctg gaaggcagag ccgacatggc cctgaagctc gtgggcgggg gccacctgat 6841 ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg 6901 cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt 6961 cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacag 7021 atgagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg 7081 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg 7141 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 7201 tcacaaataa agcatttttt tcactgccaa ctttattata catagttgat aattcactgg 7261 ccgtcgtttt acggtaccat cgatgatgat ccagacatga taagatacat tgatgagttt 7321 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct 7381 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt 7441 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc 7501 tacaaatgtg gtatggctga ttatgatcct ctagatcaga tctgcgaaga tacggccacg 7561 ggtgctcttg atcctgtggc tgattttgga ctgtgctgct cgcagctgct gatgaatcac 7621 atacttcctc cattttcttc cactgattga ctgttataat ttccctaatt tccaggtcaa 7681 ggtgctgtgc attgtggtaa tagatgtgac atgacgtcac ttccaaagga ccaatgaaca 7741 tgtctgacca atttcatata atgtgaaaac gattttcata ggcagaataa ataacattta 7801 aattaaactg ggcatcagcg caattcaatt ggtttggtaa tagcaaggga aaatagaatg 7861 aagtgatctc caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag 7921 tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta 7981 ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac 8041 tcaagtactt tacacctctg gttcttgacc ccctaccttc agcaagccca gcagatccac 8101 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt 8161 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 8221 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 8281 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 8341 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 8401 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 8461 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 8521 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 8581 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 8641 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 8701 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 8761 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 8821 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 8881 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 8941 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 9001 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 9061 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 9121 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 9181 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 9241 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 9301 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 9361 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 9421 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 9481 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 9541 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 9601 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 9661 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 9721 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 9781 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 9841 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 9901 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 9961 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 10021 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 10081 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 10141 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 10201 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 10261 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 10321 cccgaaaagt g //