LOCUS Exported 9594 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab14. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab14 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9594) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9594) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9594 /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" regulatory 5117..5126 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5123..5818 /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(5820..5844) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(6491..6511) /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(6519..6535) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 6551..6685 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6575..6594) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6629..6648 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(7357..7373) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7357..7373) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(7410..7430) /label=T3 /note="T3 promoter, forward primer" promoter complement(7410..7428) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7449..7465) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7462..7484) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7473..7489 /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(7497..7527) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7542..7563 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7680..7697) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7851..8439) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7931..7950) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8610..9470) /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 9233..9252 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9471..9575) /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 ccatggtgag cgagctgatt aaggagaaca tgcacatgaa 5161 gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg 5221 caagccctac gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc 5281 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca 5341 cacccagggc atccccgact tctttaagca gtccttcccc gagggcttca catgggagag 5401 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga 5461 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt 5521 gatgcagaag aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg 5581 cggcctggaa ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg 5641 caacttgaag accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt 5701 ctactatgtg gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga 5761 gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagaga 5821 cccagctttc ttgtacaaag tggtgacgac cgcaccatat aactattcct atattttcaa 5881 atatatcatt attggtgaca tgggggttgg caagtcctgt ttactccacc agttcacaga 5941 aaagaaattt atggctgact gtcctcatac aatcggtgtt gagtttggca cacggataat 6001 tgaagtgagt gggcagaagg tgaagcttca gatttgggac acggcaggtc aggagaggtt 6061 tcgtgctgtc acacgaagct actacagagg agccgctggg gccctcatgg tgtatgacat 6121 caccaggcga agcacgtaca atcaccttag cagctggctg actgatgcca ggaatctcac 6181 caaccccaat actgtcatca ttctaatcgg taacaaagca gacctagaag cccagcgtga 6241 tgtcacatat gaagaggcca agcagtttgc tgaagagaac ggtttgttgt tccttgaggc 6301 aagtgcaaaa acaggcgaga atgtggagga tgcttttctg gaagctgcca agaagatcta 6361 ccagaacatt caggacggca gcctggacct gaacgcagcc gagtcagggg tccagcacaa 6421 gcccaccgcc ccgcagggag ggcggctcag cagcgacgca cagccccaga aagagggctg 6481 cagctgttaa caactttatt atacatagtt gataattcac tggccgtcgt tttacggtac 6541 catcgatgat gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag 6601 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 6661 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 6721 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc 6781 tgattatgat cctctagatc agatctgcga agatacggcc acgggtgctc ttgatcctgt 6841 ggctgatttt ggactgtgct gctcgcagct gctgatgaat cacatacttc ctccattttc 6901 ttccactgat tgactgttat aatttcccta atttccaggt caaggtgctg tgcattgtgg 6961 taatagatgt gacatgacgt cacttccaaa ggaccaatga acatgtctga ccaatttcat 7021 ataatgtgaa aacgattttc ataggcagaa taaataacat ttaaattaaa ctgggcatca 7081 gcgcaattca attggtttgg taatagcaag ggaaaataga atgaagtgat ctccaaaaaa 7141 taagtacttt ttgactgtaa ataaaattgt aaggagtaaa aagtactttt ttttctaaaa 7201 aaatgtaatt aagtaaaagt aaaagtattg atttttaatt gtactcaagt aaagtaaaaa 7261 tccccaaaaa taatacttaa gtacagtaat caagtaaaat tactcaagta ctttacacct 7321 ctggttcttg accccctacc ttcagcaagc ccagcagatc cactagttct agagcggccg 7381 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg 7441 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 7501 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 7561 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 7621 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 7681 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 7741 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 7801 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 7861 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 7921 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 7981 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 8041 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 8101 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 8161 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 8221 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 8281 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 8341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 8401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 8461 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 8521 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 8581 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 8641 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 8701 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 8761 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 8821 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 8881 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 8941 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 9001 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 9061 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 9121 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 9181 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 9241 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 9301 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 9361 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 9421 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 9481 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 9541 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtg //