LOCUS Exported 9543 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab7b. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab7b SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9543) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9543) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9543 /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(1367..1387) /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(1395..1411) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1427..1561 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1451..1470) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1505..1524 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2233..2249) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2233..2249) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2286..2306) /label=T3 /note="T3 promoter, forward primer" promoter complement(2286..2304) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2325..2341) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2338..2360) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2349..2365 /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(2373..2403) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2418..2439 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2556..2573) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2727..3315) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2807..2826) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3486..4346) /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 4109..4128 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4347..4451) /gene="bla" /label=AmpR promoter primer_bind complement(4614..4635) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5531..5547 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5567..5587 /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 ggtggcttcc cgtaaaaagg tcctcctcaa 781 agtgatcatc ctaggggatt ctggagttgg gaagacctct ttgatgaacc agtatgtgaa 841 taataagttc agtaatcagt ataaggccac cattggtgca gactttctga caaaggaggt 901 gatggtggac gacagactgg tgaccatgca gatttgggac acagccggtc aggagcggtt 961 tcagtctctg ggcgtggcct tctacagagg ggcggactgt tgtgtgctgg tgtatgatgt 1021 cacggcgccc accacgttta agactctgga cagctggagg gacgagtttc tcattcaggc 1081 cagtacaagc gacccggaga acttcccctt tgttgtgctg ggcaacaaaa tcgacctgga 1141 caaccgacag gtatcgacaa agcgagcaca agcgtggtgc cagagtaaga acaacattcc 1201 ctattttgag accagcgcca aagaagccat taatgtggaa caggccttcc agaccatcgc 1261 tcgcaatgcg cttaaacagg agtcggtgga caaatatgat ttcccggacg agatcaaact 1321 aggaaacgac agacccatgt ctaacggtga aggctgcagc tgctgacaac tttattatac 1381 atagttgata attcactggc cgtcgtttta cggtaccatc gatgatgatc cagacatgat 1441 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat 1501 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt 1561 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt 1621 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcctc tagatcagat 1681 ctgcgaagat acggccacgg gtgctcttga tcctgtggct gattttggac tgtgctgctc 1741 gcagctgctg atgaatcaca tacttcctcc attttcttcc actgattgac tgttataatt 1801 tccctaattt ccaggtcaag gtgctgtgca ttgtggtaat agatgtgaca tgacgtcact 1861 tccaaaggac caatgaacat gtctgaccaa tttcatataa tgtgaaaacg attttcatag 1921 gcagaataaa taacatttaa attaaactgg gcatcagcgc aattcaattg gtttggtaat 1981 agcaagggaa aatagaatga agtgatctcc aaaaaataag tactttttga ctgtaaataa 2041 aattgtaagg agtaaaaagt actttttttt ctaaaaaaat gtaattaagt aaaagtaaaa 2101 gtattgattt ttaattgtac tcaagtaaag taaaaatccc caaaaataat acttaagtac 2161 agtaatcaag taaaattact caagtacttt acacctctgg ttcttgaccc cctaccttca 2221 gcaagcccag cagatccact agttctagag cggccgccac cgcggtggag ctccagcttt 2281 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct 2341 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 2401 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 2461 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 2521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 2581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 2641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 2701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 2761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 2821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 2881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 2941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 3001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 3061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 3121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 3181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 3241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 3301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 3361 gaaaactcac gttaaggcat tttggtcatg agattatcaa aaaggatctt cacctagatc 3421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 3481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 3541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 3601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 3661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 3721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 3781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 3841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 3901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 3961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 4021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 4081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 4141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 4201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 4261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 4321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 4381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 4441 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt 4501 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 4561 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 4621 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 4681 atcagggcga tggcccacgc tggctaagaa ctcatcagcc tccccggtcc atctacccac 4741 gtaccaatgc accaattggc cacaatgacg gctactacat ggtgccattc cttcctcttt 4801 ataggaatgg agactacctc ctgtccaaca atgctcttgg atacgagtac gcctacctgt 4861 tggacccagg tcattgcaca acaccagaaa tgccctctga tctgcaaaag acgtgaatat 4921 ctgttcagac acccatatcc actctgttcc acacaggtca gaggtttgtc caggagttct 4981 tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt gaaagtacaa gtacttaggg 5041 aaaattttac tcaattaaaa gtaaaagtat ctggctagaa tcttacttga gtaaaagtaa 5101 aaaagtactc cattaaaatt gtacttgagt attaaggaag taaaagtaaa agcaagaaag 5161 aaaactagag attcttgttt aagcttttaa tctcaaaaaa cattaaatga aatgcataca 5221 aggttttatc ctgctttaga actgtttgta tttaattatc aaactataag acagacaatc 5281 taatgccagt acacgctact caaagttgta aaacctcaga tttaacttca gtagaagctg 5341 attctcaaaa ttgttagtgt caagcctagc tcttttgggg ctgaaaagca atcctgcagt 5401 gctgaaaagc ctctcacagg cagccgatgc gggaagaggt gtattagtct tgatagagag 5461 gctgcaaata gcaggaaacg tgagcagaga ctccctggtg tctgaaacac aggccagatg 5521 ggccctcgag caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga 5581 aaagttggca cgcgcagatt ccgtgtgggc ctggtaatat cttaccattt taagctattt 5641 taagattagg gaaaaaaatt aaataaaact actttaaaat tactttcctt aaaaagcaac 5701 atatttattt agtttttgtt taactggctc aagattgtaa tattttactt ttacatgttt 5761 ttttaagtac ctaaatgtgt tcaaaagtaa agtatgttat tgtttttttt ttttaatgtt 5821 agctaagctt taaattgtaa tatattactt ttaaaagtga ctttttaaaa atgtaaataa 5881 aaaagtaact ataaatgtgt aatgcatgtt atttctgtgg gtttttaatt ttcatctcaa 5941 ctgggcttac aattttaaca taataaaaat caaatcaaat caaatcaggc ccttatttgt 6001 cacatacact cttgtatact gaaataaact aaatgttgag aaaaatttaa atacaccaaa 6061 ataacgcata ccttttttgt aattacctaa gtaaaacatt tagttccttt tttaagtgac 6121 tcaagattgt aatctattaa ttgtaaaaga taaaactagt gatcaatctg acgtgcttct 6181 atgcagcctg tgagattaat tgtgatgttt aattattaaa caatttaaat ggatcaaaca 6241 tttcacatgt gcttattgtg tccaacaaaa ccagcagaag cacttaaaaa tattgcatga 6301 caacatgagt ttttaatgta aattatatat acttcctaaa cgaagttaat catatttcta 6361 aaataataat gtcattgaaa tatttaaata aatgataagc tttagttgat tcaacataaa 6421 attgaaagca ataaggactt ttttttttta cagcggtgaa gcaaaagcat gtaataatca 6481 tataatataa taaataaata aataaataat ggctcttggt gcaaatatga acactttaaa 6541 taaatataga ataaatatga tcaataagtc ataacgatgt gttttatgtt gattataatt 6601 taaacatgtt tctaacgcaa gttttaaaaa ctgttttaaa gtattatttt aagttataag 6661 atcaatttga ctcaacattt tgtagtgaaa atatgttgtg taaaattaat ataatataca 6721 ctataaaaat gcagggttcc acacaataca ttaatgttgt cccaacacaa atcgattaag 6781 ttaacaaata tatgtggatt gaacataaaa aattaagttg tcccaatgaa atctcaagaa 6841 ttgtgttgtt tcagctcatt ttaaataagt agttttaaca agtaaaaaat aacatatttt 6901 tgagtgtgta tatatatata ttacaaaata taataatata aaatgtaaaa gatataaacc 6961 atgctacata tatttaagag ctattttgca cattactttt ttgcaagcca ttttagtgtt 7021 ttttttacaa atatatatta gctaaatata caatatctta aatatacaat taatcagaca 7081 tgcaattaat gcatttaacc tacataaaaa tacatgatca acatatgttt tatgctgatt 7141 acaactaaat caaacgtaat gaaattaatt tataaaagcg atttaaatat gtcaatattt 7201 taagttaaaa tcacttgaaa cctactataa atgaacttaa attaaaaaaa tctcctttaa 7261 ttccaatgca acagtgttgt tttttcagct gaggaccccc ggtgcagatg gttggccccc 7321 ggcccgctat tctaaggaag cctttcttaa ttttagggcg gcgctgcatg cgctcagatc 7381 agattcacgg gtggagaggg ttattttctc ttttttattt tttcttgcct gctcatattt 7441 gggcctgtgt gaaccccagg ctccgcctcg gcccccagga gccgctctta tttaaagggg 7501 atcctcgcct tcagtcttcg tcccagtgga agcatcagtc ttgtgataac aacttggtga 7561 gtcctacaga ccagcgctcg actctaggct ttaaatgcat tgatttctct ggattagcta 7621 ccgggagagt tttaactttg cttattagtc ttgtgctcga actgctaggt ttaaagagtg 7681 ttttctctaa tgcatgcatg aagcgacagc tgaaaaccag aaatagagcg tttgtcctcc 7741 tgttaaactc tagcctttct ttggcatttg tcagtccgag tcttcagctg ccggcaggag 7801 attgggtttc agtaacactt gctgagcaca aactacacgt gctttcttca gatttaattc 7861 attattaatc attctagatt taaatgatgt tcttactcat tgctgattat tttagccttg 7921 cggagcagat taaatgagca cgtgtttcat taagattgat ttaaatataa ttaatatttc 7981 accactgtag ttttaaaatt gccttctaag tcatggttga tttttaatag gtatcggctc 8041 taattagatt actattttgg tttttaaatg ttatttctat ttcatttgta ggctcgtttt 8101 taatgtgatt ttattagatt ctagttttaa aatcatctct tagccttaat gagcagactg 8161 aatgaacatg tttcattaag attaacttca ttataatgaa ttatttaatc attctagtgg 8221 gttataaatc agctctattt aaattaattg tcgtgtttat ttctatttaa tttttgatta 8281 ttttttcagg gtcgttttta tgtgattttt tcccgtttct aattttaaac taattgtttg 8341 atttttatat gattcgactc tttttaaatg aattgttttg ctttttttat tgagcatata 8401 tatattatac aatttcaatc aacaattgtt gtgcttttta aatgttgttt ctatttcatt 8461 tctagggtcg attttattcg attctagttt taaaatcgtg atttttaact tctaacctta 8521 ctgagcagac tgaatgaaca tgtttcatca agattaactt cattatagtt aattattgaa 8581 tcgtgctagt ggtttataaa tcagctctat ttaaattaat catcatcatt atttctattt 8641 aatttttatt atttctttag gattattttt atgtgatttt tttcacgttc tagttttaaa 8701 ctcaatgctc attactgctc acccttgctg agcagtcttt cgttaagatt aaattaatta 8761 ataaccattc tagttttaaa acagtgtctt aaggcatagc tgatatttta caagtttact 8821 atttaatata tctcctttct ttttttaggg tcgatttcat ctgatttgat taatttttat 8881 tatcttcact aagctttttg ttctaggttt tattctcagt attattactg ttatatttga 8941 actgaatttc tcctcatagt ttattcattt tatttattta ttttttgttt attcgctatt 9001 ttgtatttta gtttttaatt agaaatttta ttagaatcgt tttgaactag tttttatgca 9061 ctctaacatt aatattaaat cataactatt atttttatta taatttattt gtgtttatct 9121 ttttgtgtag ctttcatgtg tttttatttt agttttatta tgtttgcatt gaattttttg 9181 gactagttta tataacgata tacagaaaag ccatgacaac aaatgttttt ataataccta 9241 ttaagtataa gctaatcaga tttccactat tctgttcaaa gttaacatta ttagccattc 9301 tgatggatca tttgacattt gtcatatgtc tttatgaaca ctgctctttc agatgccact 9361 tttaaagctg ttttttacag taaatgaacg tttattatta tgacattatt tcctaaggcc 9421 tctagagtac agcaggatgt ggcgtgtgca ttaccgtgta attatgctga ctgtaaatgt 9481 gcatttatag actgtcagca tgtctgactg tgcgctgttg tgattggtcc tgcacagacc 9541 aag //