LOCUS Exported 9578 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab5c. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab6a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9578) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9578) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9578 /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(6475..6495) /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(6503..6519) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 6535..6669 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6559..6578) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6613..6632 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(7341..7357) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7341..7357) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(7394..7414) /label=T3 /note="T3 promoter, forward primer" promoter complement(7394..7412) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7433..7449) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7446..7468) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7457..7473 /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(7481..7511) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7526..7547 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7664..7681) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7835..8423) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7915..7934) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8594..9454) /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 9217..9236 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9455..9559) /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 tggtgtctgc agcaggagat ttcggcaacc cgctccgcaa 5881 attcaagcta gtttttctgg gagagcagag cgtgggaaag acgtcgttga ttaccagatt 5941 tatgtacgac agtttcgata acacctacca ggccacgata ggaattgatt tcttgtcgaa 6001 aaccatgtgt cttgaggaca gaacaatcag gctgcagctt tgggacacag caggacagga 6061 gcgtttccgt agcctcattc ccagttacat ccgagattca gccgctgctg ttgtagttta 6121 cgacatcaca aatgtcaact cattccagca gaccaccaaa tggatcgatg acgtccgaac 6181 ggaaagagga agtgatgtca tcatcatgct agtgggaaac aaaacagatc ttgcagacaa 6241 aaggcaagtg tctatcgagg aaggagaaag gaaagccaaa gaactgaatg taatgtttat 6301 tgaaactagt gctaaagccg gatacaacgt caagcagctt ttccgccgtg tagcagcagc 6361 ccttcctggt atggagagca cacaagacaa gagcagagag gacatgatcg acataaaatt 6421 ggaaaagcca ccagagcagc cggtcagcga aggcggctgt tcatgctgag gcttcaactt 6481 tattatacat agttgataat tcactggccg tcgttttacg gtaccatcga tgatgatcca 6541 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa 6601 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 6661 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg 6721 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcctcta 6781 gatcagatct gcgaagatac ggccacgggt gctcttgatc ctgtggctga ttttggactg 6841 tgctgctcgc agctgctgat gaatcacata cttcctccat tttcttccac tgattgactg 6901 ttataatttc cctaatttcc aggtcaaggt gctgtgcatt gtggtaatag atgtgacatg 6961 acgtcacttc caaaggacca atgaacatgt ctgaccaatt tcatataatg tgaaaacgat 7021 tttcataggc agaataaata acatttaaat taaactgggc atcagcgcaa ttcaattggt 7081 ttggtaatag caagggaaaa tagaatgaag tgatctccaa aaaataagta ctttttgact 7141 gtaaataaaa ttgtaaggag taaaaagtac ttttttttct aaaaaaatgt aattaagtaa 7201 aagtaaaagt attgattttt aattgtactc aagtaaagta aaaatcccca aaaataatac 7261 ttaagtacag taatcaagta aaattactca agtactttac acctctggtt cttgaccccc 7321 taccttcagc aagcccagca gatccactag ttctagagcg gccgccaccg cggtggagct 7381 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc 7441 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7501 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7561 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7621 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7681 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 7741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 8041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 8101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 8161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 8221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 8281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 8341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 8401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8521 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8581 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 8701 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 8761 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 8821 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 8881 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 8941 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 9001 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 9061 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 9121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 9181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 9241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 9301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 9361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 9421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9541 aacaaatagg ggttccgcgc acatttcccc gaaaagtg //