LOCUS Exported 9562 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab7a. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab7a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9562) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9562) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9562 /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(1386..1406) /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(1414..1430) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1446..1580 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1470..1489) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1524..1543 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2252..2268) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2252..2268) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2305..2325) /label=T3 /note="T3 promoter, forward primer" promoter complement(2305..2323) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2344..2360) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2357..2379) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2368..2384 /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(2392..2422) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2437..2458 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2575..2592) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2746..3334) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2826..2845) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3505..4365) /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 4128..4147 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4366..4470) /gene="bla" /label=AmpR promoter primer_bind complement(4633..4654) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5550..5566 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5586..5606 /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 gggcgccacc atgacatcaa ggaagaaagt 781 tcttctgaag gtgatcattc ttggagattc cggggttgga aaaacatctc taatgaacca 841 gtatgtgaat aaaaagttca gtaatcagta taaagccact ataggagcag atttcctgac 901 aaaagaagtg atggttgacg accggcttgt cacaatgcag atatgggaca cagcaggtca 961 ggagcggttt cagtctctgg gtgttgcttt ctatcgtggt gctgactgct gcgtcctggt 1021 gtttgacgtc actgccccga acaccttcaa gacactggac agctggaggg atgagtttct 1081 gatccaggcc agtcctcgag acccagaaaa cttccctttc gtggtgctgg gcaacaaaat 1141 cgatctggag aatagacagg taacaactaa gcgagcacaa gcttggtgtc agagtaagaa 1201 caacatccca tattttgaga ccagtgcaaa ggaggccatc aacgtagagc aggcattcca 1261 gaccatcgcg cgcaatgcac tcaaacagga aactgaagtg gagttataca acgagttccc 1321 cgagccaatt aaactagacc ggaacgatag agccaaacca tcagcagaga cctgtagctg 1381 ctgaccaact ttattataca tagttgataa ttcactggcc gtcgttttac ggtaccatcg 1441 atgatgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 1501 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 1561 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 1621 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt 1681 atgatcctct agatcagatc tgcgaagata cggccacggg tgctcttgat cctgtggctg 1741 attttggact gtgctgctcg cagctgctga tgaatcacat acttcctcca ttttcttcca 1801 ctgattgact gttataattt ccctaatttc caggtcaagg tgctgtgcat tgtggtaata 1861 gatgtgacat gacgtcactt ccaaaggacc aatgaacatg tctgaccaat ttcatataat 1921 gtgaaaacga ttttcatagg cagaataaat aacatttaaa ttaaactggg catcagcgca 1981 attcaattgg tttggtaata gcaagggaaa atagaatgaa gtgatctcca aaaaataagt 2041 actttttgac tgtaaataaa attgtaagga gtaaaaagta cttttttttc taaaaaaatg 2101 taattaagta aaagtaaaag tattgatttt taattgtact caagtaaagt aaaaatcccc 2161 aaaaataata cttaagtaca gtaatcaagt aaaattactc aagtacttta cacctctggt 2221 tcttgacccc ctaccttcag caagcccagc agatccacta gttctagagc ggccgccacc 2281 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc 2341 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 2401 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 2461 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 2521 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 2581 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 2641 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 2701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 2761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 2821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 2881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 2941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 3001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 3061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 3121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 3181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 3241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 3301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 3361 gtctgacgct cagtggaacg aaaactcacg ttaaggcatt ttggtcatga gattatcaaa 3421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 3481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 3541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 3601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 3661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 3721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 3781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 3841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 3901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 3961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 4021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 4081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 4141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 4201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 4261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 4321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 4381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 4441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt 4501 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 4561 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 4621 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 4681 aaagggcgaa aaaccgtcta tcagggcgat ggcccacgct ggctaagaac tcatcagcct 4741 ccccggtcca tctacccacg taccaatgca ccaattggcc acaatgacgg ctactacatg 4801 gtgccattcc ttcctcttta taggaatgga gactacctcc tgtccaacaa tgctcttgga 4861 tacgagtacg cctacctgtt ggacccaggt cattgcacaa caccagaaat gccctctgat 4921 ctgcaaaaga cgtgaatatc tgttcagaca cccatatcca ctctgttcca cacaggtcag 4981 aggtttgtcc aggagttctt gacagaggtg taaaaagtac tcaaaaattt tactcaagtg 5041 aaagtacaag tacttaggga aaattttact caattaaaag taaaagtatc tggctagaat 5101 cttacttgag taaaagtaaa aaagtactcc attaaaattg tacttgagta ttaaggaagt 5161 aaaagtaaaa gcaagaaaga aaactagaga ttcttgttta agcttttaat ctcaaaaaac 5221 attaaatgaa atgcatacaa ggttttatcc tgctttagaa ctgtttgtat ttaattatca 5281 aactataaga cagacaatct aatgccagta cacgctactc aaagttgtaa aacctcagat 5341 ttaacttcag tagaagctga ttctcaaaat tgttagtgtc aagcctagct cttttggggc 5401 tgaaaagcaa tcctgcagtg ctgaaaagcc tctcacaggc agccgatgcg ggaagaggtg 5461 tattagtctt gatagagagg ctgcaaatag caggaaacgt gagcagagac tccctggtgt 5521 ctgaaacaca ggccagatgg gccctcgagc aggaaacagc tatgaccatg attacgccaa 5581 gctatcaact ttgtatagaa aagttggcac gcgcagattc cgtgtgggcc tggtaatatc 5641 ttaccatttt aagctatttt aagattaggg aaaaaaatta aataaaacta ctttaaaatt 5701 actttcctta aaaagcaaca tatttattta gtttttgttt aactggctca agattgtaat 5761 attttacttt tacatgtttt tttaagtacc taaatgtgtt caaaagtaaa gtatgttatt 5821 gttttttttt tttaatgtta gctaagcttt aaattgtaat atattacttt taaaagtgac 5881 tttttaaaaa tgtaaataaa aaagtaacta taaatgtgta atgcatgtta tttctgtggg 5941 tttttaattt tcatctcaac tgggcttaca attttaacat aataaaaatc aaatcaaatc 6001 aaatcaggcc cttatttgtc acatacactc ttgtatactg aaataaacta aatgttgaga 6061 aaaatttaaa tacaccaaaa taacgcatac cttttttgta attacctaag taaaacattt 6121 agttcctttt ttaagtgact caagattgta atctattaat tgtaaaagat aaaactagtg 6181 atcaatctga cgtgcttcta tgcagcctgt gagattaatt gtgatgttta attattaaac 6241 aatttaaatg gatcaaacat ttcacatgtg cttattgtgt ccaacaaaac cagcagaagc 6301 acttaaaaat attgcatgac aacatgagtt tttaatgtaa attatatata cttcctaaac 6361 gaagttaatc atatttctaa aataataatg tcattgaaat atttaaataa atgataagct 6421 ttagttgatt caacataaaa ttgaaagcaa taaggacttt ttttttttac agcggtgaag 6481 caaaagcatg taataatcat ataatataat aaataaataa ataaataatg gctcttggtg 6541 caaatatgaa cactttaaat aaatatagaa taaatatgat caataagtca taacgatgtg 6601 ttttatgttg attataattt aaacatgttt ctaacgcaag ttttaaaaac tgttttaaag 6661 tattatttta agttataaga tcaatttgac tcaacatttt gtagtgaaaa tatgttgtgt 6721 aaaattaata taatatacac tataaaaatg cagggttcca cacaatacat taatgttgtc 6781 ccaacacaaa tcgattaagt taacaaatat atgtggattg aacataaaaa attaagttgt 6841 cccaatgaaa tctcaagaat tgtgttgttt cagctcattt taaataagta gttttaacaa 6901 gtaaaaaata acatattttt gagtgtgtat atatatatat tacaaaatat aataatataa 6961 aatgtaaaag atataaacca tgctacatat atttaagagc tattttgcac attacttttt 7021 tgcaagccat tttagtgttt tttttacaaa tatatattag ctaaatatac aatatcttaa 7081 atatacaatt aatcagacat gcaattaatg catttaacct acataaaaat acatgatcaa 7141 catatgtttt atgctgatta caactaaatc aaacgtaatg aaattaattt ataaaagcga 7201 tttaaatatg tcaatatttt aagttaaaat cacttgaaac ctactataaa tgaacttaaa 7261 ttaaaaaaat ctcctttaat tccaatgcaa cagtgttgtt ttttcagctg aggacccccg 7321 gtgcagatgg ttggcccccg gcccgctatt ctaaggaagc ctttcttaat tttagggcgg 7381 cgctgcatgc gctcagatca gattcacggg tggagagggt tattttctct tttttatttt 7441 ttcttgcctg ctcatatttg ggcctgtgtg aaccccaggc tccgcctcgg cccccaggag 7501 ccgctcttat ttaaagggga tcctcgcctt cagtcttcgt cccagtggaa gcatcagtct 7561 tgtgataaca acttggtgag tcctacagac cagcgctcga ctctaggctt taaatgcatt 7621 gatttctctg gattagctac cgggagagtt ttaactttgc ttattagtct tgtgctcgaa 7681 ctgctaggtt taaagagtgt tttctctaat gcatgcatga agcgacagct gaaaaccaga 7741 aatagagcgt ttgtcctcct gttaaactct agcctttctt tggcatttgt cagtccgagt 7801 cttcagctgc cggcaggaga ttgggtttca gtaacacttg ctgagcacaa actacacgtg 7861 ctttcttcag atttaattca ttattaatca ttctagattt aaatgatgtt cttactcatt 7921 gctgattatt ttagccttgc ggagcagatt aaatgagcac gtgtttcatt aagattgatt 7981 taaatataat taatatttca ccactgtagt tttaaaattg ccttctaagt catggttgat 8041 ttttaatagg tatcggctct aattagatta ctattttggt ttttaaatgt tatttctatt 8101 tcatttgtag gctcgttttt aatgtgattt tattagattc tagttttaaa atcatctctt 8161 agccttaatg agcagactga atgaacatgt ttcattaaga ttaacttcat tataatgaat 8221 tatttaatca ttctagtggg ttataaatca gctctattta aattaattgt cgtgtttatt 8281 tctatttaat ttttgattat tttttcaggg tcgtttttat gtgatttttt cccgtttcta 8341 attttaaact aattgtttga tttttatatg attcgactct ttttaaatga attgttttgc 8401 tttttttatt gagcatatat atattataca atttcaatca acaattgttg tgctttttaa 8461 atgttgtttc tatttcattt ctagggtcga ttttattcga ttctagtttt aaaatcgtga 8521 tttttaactt ctaaccttac tgagcagact gaatgaacat gtttcatcaa gattaacttc 8581 attatagtta attattgaat cgtgctagtg gtttataaat cagctctatt taaattaatc 8641 atcatcatta tttctattta atttttatta tttctttagg attattttta tgtgattttt 8701 ttcacgttct agttttaaac tcaatgctca ttactgctca cccttgctga gcagtctttc 8761 gttaagatta aattaattaa taaccattct agttttaaaa cagtgtctta aggcatagct 8821 gatattttac aagtttacta tttaatatat ctcctttctt tttttagggt cgatttcatc 8881 tgatttgatt aatttttatt atcttcacta agctttttgt tctaggtttt attctcagta 8941 ttattactgt tatatttgaa ctgaatttct cctcatagtt tattcatttt atttatttat 9001 tttttgttta ttcgctattt tgtattttag tttttaatta gaaattttat tagaatcgtt 9061 ttgaactagt ttttatgcac tctaacatta atattaaatc ataactatta tttttattat 9121 aatttatttg tgtttatctt tttgtgtagc tttcatgtgt ttttatttta gttttattat 9181 gtttgcattg aattttttgg actagtttat ataacgatat acagaaaagc catgacaaca 9241 aatgttttta taatacctat taagtataag ctaatcagat ttccactatt ctgttcaaag 9301 ttaacattat tagccattct gatggatcat ttgacatttg tcatatgtct ttatgaacac 9361 tgctctttca gatgccactt ttaaagctgt tttttacagt aaatgaacgt ttattattat 9421 gacattattt cctaaggcct ctagagtaca gcaggatgtg gcgtgtgcat taccgtgtaa 9481 ttatgctgac tgtaaatgtg catttataga ctgtcagcat gtctgactgt gcgctgttgt 9541 gattggtcct gcacagacca ag //