LOCUS Exported 9597 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab27a. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab27a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9597) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9597) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9597 /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(1421..1441) /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(1449..1465) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1481..1615 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1505..1524) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1559..1578 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2287..2303) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2287..2303) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2340..2360) /label=T3 /note="T3 promoter, forward primer" promoter complement(2340..2358) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2379..2395) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2392..2414) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2403..2419 /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(2427..2457) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2472..2493 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2610..2627) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2781..3369) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2861..2880) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3540..4400) /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 4163..4182 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4401..4505) /gene="bla" /label=AmpR promoter primer_bind complement(4668..4689) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5585..5601 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5621..5641 /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 ggtgtccgat ggggactatg attacctcat 781 caagttcctc gctctggggg actctggggt gggaaagacc agcttcctat atcagtacac 841 agacagcaaa ttcaactcca aattcataac cacagtggga atagacttca gagaaaaacg 901 agtggtgtat aaatccagtg gtcctgatgg cacagcaggc agaggccaga ggatccacat 961 ccagctttgg gatacggccg gacaggagag gtttcgaagt ttgaccactg ctttcttcag 1021 agatgcgatg ggctttctgc tgctctttga cctcaccaac gagcagagct tcctgaatgt 1081 gcgcaactgg atgagtcagc tgcagacaca tgcgtactgt gagaatcctg atatcgtctt 1141 gtgtggaaat aaatcagacc tggaggagca gcgtgccgtc aaagcggaga acgcaagaga 1201 actggcagag aaatacggcg tgccatactt cgagacgagc gcggcgaacg gtgagaatgt 1261 gagtcgtgcg gtggaggttc tgctggatct gatcatgaag cgcatggaga gatgtgtgga 1321 taaatcctgg atccccgacg ggacagtccg cagcaacggc cacagcaccg ccgacctgac 1381 agaaccacac gaccccgagc agagcaagtg tgcatgctaa caactttatt atacatagtt 1441 gataattcac tggccgtcgt tttacggtac catcgatgat gatccagaca tgataagata 1501 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga 1561 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa 1621 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag 1681 caagtaaaac ctctacaaat gtggtatggc tgattatgat cctctagatc agatctgcga 1741 agatacggcc acgggtgctc ttgatcctgt ggctgatttt ggactgtgct gctcgcagct 1801 gctgatgaat cacatacttc ctccattttc ttccactgat tgactgttat aatttcccta 1861 atttccaggt caaggtgctg tgcattgtgg taatagatgt gacatgacgt cacttccaaa 1921 ggaccaatga acatgtctga ccaatttcat ataatgtgaa aacgattttc ataggcagaa 1981 taaataacat ttaaattaaa ctgggcatca gcgcaattca attggtttgg taatagcaag 2041 ggaaaataga atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt 2101 aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg 2161 atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat 2221 caagtaaaat tactcaagta ctttacacct ctggttcttg accccctacc ttcagcaagc 2281 ccagcagatc cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc 2341 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga 2401 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 2461 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 2521 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 2581 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 2641 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 2701 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 2761 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 2821 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 2881 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 2941 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 3001 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 3061 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 3121 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 3181 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 3241 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 3301 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 3361 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 3421 tcacgttaag gcattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 3481 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 3541 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 3601 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 3661 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 3721 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 3781 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 3841 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 3901 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 3961 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 4021 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 4081 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 4141 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 4201 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 4261 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 4321 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 4381 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 4441 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 4501 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat 4561 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 4621 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 4681 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 4741 gcgatggccc acgctggcta agaactcatc agcctccccg gtccatctac ccacgtacca 4801 atgcaccaat tggccacaat gacggctact acatggtgcc attccttcct ctttatagga 4861 atggagacta cctcctgtcc aacaatgctc ttggatacga gtacgcctac ctgttggacc 4921 caggtcattg cacaacacca gaaatgccct ctgatctgca aaagacgtga atatctgttc 4981 agacacccat atccactctg ttccacacag gtcagaggtt tgtccaggag ttcttgacag 5041 aggtgtaaaa agtactcaaa aattttactc aagtgaaagt acaagtactt agggaaaatt 5101 ttactcaatt aaaagtaaaa gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt 5161 actccattaa aattgtactt gagtattaag gaagtaaaag taaaagcaag aaagaaaact 5221 agagattctt gtttaagctt ttaatctcaa aaaacattaa atgaaatgca tacaaggttt 5281 tatcctgctt tagaactgtt tgtatttaat tatcaaacta taagacagac aatctaatgc 5341 cagtacacgc tactcaaagt tgtaaaacct cagatttaac ttcagtagaa gctgattctc 5401 aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa agcaatcctg cagtgctgaa 5461 aagcctctca caggcagccg atgcgggaag aggtgtatta gtcttgatag agaggctgca 5521 aatagcagga aacgtgagca gagactccct ggtgtctgaa acacaggcca gatgggccct 5581 cgagcaggaa acagctatga ccatgattac gccaagctat caactttgta tagaaaagtt 5641 ggcacgcgca gattccgtgt gggcctggta atatcttacc attttaagct attttaagat 5701 tagggaaaaa aattaaataa aactacttta aaattacttt ccttaaaaag caacatattt 5761 atttagtttt tgtttaactg gctcaagatt gtaatatttt acttttacat gtttttttaa 5821 gtacctaaat gtgttcaaaa gtaaagtatg ttattgtttt ttttttttaa tgttagctaa 5881 gctttaaatt gtaatatatt acttttaaaa gtgacttttt aaaaatgtaa ataaaaaagt 5941 aactataaat gtgtaatgca tgttatttct gtgggttttt aattttcatc tcaactgggc 6001 ttacaatttt aacataataa aaatcaaatc aaatcaaatc aggcccttat ttgtcacata 6061 cactcttgta tactgaaata aactaaatgt tgagaaaaat ttaaatacac caaaataacg 6121 catacctttt ttgtaattac ctaagtaaaa catttagttc cttttttaag tgactcaaga 6181 ttgtaatcta ttaattgtaa aagataaaac tagtgatcaa tctgacgtgc ttctatgcag 6241 cctgtgagat taattgtgat gtttaattat taaacaattt aaatggatca aacatttcac 6301 atgtgcttat tgtgtccaac aaaaccagca gaagcactta aaaatattgc atgacaacat 6361 gagtttttaa tgtaaattat atatacttcc taaacgaagt taatcatatt tctaaaataa 6421 taatgtcatt gaaatattta aataaatgat aagctttagt tgattcaaca taaaattgaa 6481 agcaataagg actttttttt tttacagcgg tgaagcaaaa gcatgtaata atcatataat 6541 ataataaata aataaataaa taatggctct tggtgcaaat atgaacactt taaataaata 6601 tagaataaat atgatcaata agtcataacg atgtgtttta tgttgattat aatttaaaca 6661 tgtttctaac gcaagtttta aaaactgttt taaagtatta ttttaagtta taagatcaat 6721 ttgactcaac attttgtagt gaaaatatgt tgtgtaaaat taatataata tacactataa 6781 aaatgcaggg ttccacacaa tacattaatg ttgtcccaac acaaatcgat taagttaaca 6841 aatatatgtg gattgaacat aaaaaattaa gttgtcccaa tgaaatctca agaattgtgt 6901 tgtttcagct cattttaaat aagtagtttt aacaagtaaa aaataacata tttttgagtg 6961 tgtatatata tatattacaa aatataataa tataaaatgt aaaagatata aaccatgcta 7021 catatattta agagctattt tgcacattac ttttttgcaa gccattttag tgtttttttt 7081 acaaatatat attagctaaa tatacaatat cttaaatata caattaatca gacatgcaat 7141 taatgcattt aacctacata aaaatacatg atcaacatat gttttatgct gattacaact 7201 aaatcaaacg taatgaaatt aatttataaa agcgatttaa atatgtcaat attttaagtt 7261 aaaatcactt gaaacctact ataaatgaac ttaaattaaa aaaatctcct ttaattccaa 7321 tgcaacagtg ttgttttttc agctgaggac ccccggtgca gatggttggc ccccggcccg 7381 ctattctaag gaagcctttc ttaattttag ggcggcgctg catgcgctca gatcagattc 7441 acgggtggag agggttattt tctctttttt attttttctt gcctgctcat atttgggcct 7501 gtgtgaaccc caggctccgc ctcggccccc aggagccgct cttatttaaa ggggatcctc 7561 gccttcagtc ttcgtcccag tggaagcatc agtcttgtga taacaacttg gtgagtccta 7621 cagaccagcg ctcgactcta ggctttaaat gcattgattt ctctggatta gctaccggga 7681 gagttttaac tttgcttatt agtcttgtgc tcgaactgct aggtttaaag agtgttttct 7741 ctaatgcatg catgaagcga cagctgaaaa ccagaaatag agcgtttgtc ctcctgttaa 7801 actctagcct ttctttggca tttgtcagtc cgagtcttca gctgccggca ggagattggg 7861 tttcagtaac acttgctgag cacaaactac acgtgctttc ttcagattta attcattatt 7921 aatcattcta gatttaaatg atgttcttac tcattgctga ttattttagc cttgcggagc 7981 agattaaatg agcacgtgtt tcattaagat tgatttaaat ataattaata tttcaccact 8041 gtagttttaa aattgccttc taagtcatgg ttgattttta ataggtatcg gctctaatta 8101 gattactatt ttggttttta aatgttattt ctatttcatt tgtaggctcg tttttaatgt 8161 gattttatta gattctagtt ttaaaatcat ctcttagcct taatgagcag actgaatgaa 8221 catgtttcat taagattaac ttcattataa tgaattattt aatcattcta gtgggttata 8281 aatcagctct atttaaatta attgtcgtgt ttatttctat ttaatttttg attatttttt 8341 cagggtcgtt tttatgtgat tttttcccgt ttctaatttt aaactaattg tttgattttt 8401 atatgattcg actcttttta aatgaattgt tttgcttttt ttattgagca tatatatatt 8461 atacaatttc aatcaacaat tgttgtgctt tttaaatgtt gtttctattt catttctagg 8521 gtcgatttta ttcgattcta gttttaaaat cgtgattttt aacttctaac cttactgagc 8581 agactgaatg aacatgtttc atcaagatta acttcattat agttaattat tgaatcgtgc 8641 tagtggttta taaatcagct ctatttaaat taatcatcat cattatttct atttaatttt 8701 tattatttct ttaggattat ttttatgtga tttttttcac gttctagttt taaactcaat 8761 gctcattact gctcaccctt gctgagcagt ctttcgttaa gattaaatta attaataacc 8821 attctagttt taaaacagtg tcttaaggca tagctgatat tttacaagtt tactatttaa 8881 tatatctcct ttcttttttt agggtcgatt tcatctgatt tgattaattt ttattatctt 8941 cactaagctt tttgttctag gttttattct cagtattatt actgttatat ttgaactgaa 9001 tttctcctca tagtttattc attttattta tttatttttt gtttattcgc tattttgtat 9061 tttagttttt aattagaaat tttattagaa tcgttttgaa ctagttttta tgcactctaa 9121 cattaatatt aaatcataac tattattttt attataattt atttgtgttt atctttttgt 9181 gtagctttca tgtgttttta ttttagtttt attatgtttg cattgaattt tttggactag 9241 tttatataac gatatacaga aaagccatga caacaaatgt ttttataata cctattaagt 9301 ataagctaat cagatttcca ctattctgtt caaagttaac attattagcc attctgatgg 9361 atcatttgac atttgtcata tgtctttatg aacactgctc tttcagatgc cacttttaaa 9421 gctgtttttt acagtaaatg aacgtttatt attatgacat tatttcctaa ggcctctaga 9481 gtacagcagg atgtggcgtg tgcattaccg tgtaattatg ctgactgtaa atgtgcattt 9541 atagactgtc agcatgtctg actgtgcgct gttgtgattg gtcctgcaca gaccaag //