LOCUS Exported 9596 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab3ab. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab3c SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9596) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9596) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9596 /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(1420..1440) /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(1448..1464) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1480..1614 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1504..1523) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1558..1577 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2286..2302) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2286..2302) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2339..2359) /label=T3 /note="T3 promoter, forward primer" promoter complement(2339..2357) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2378..2394) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2391..2413) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2402..2418 /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(2426..2456) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2471..2492 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2609..2626) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2780..3368) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2860..2879) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3539..4399) /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 4162..4181 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4400..4504) /gene="bla" /label=AmpR promoter primer_bind complement(4667..4688) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5584..5600 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5620..5640 /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 ggtggctgct acgcaagaca acaaacaaaa 781 ggagaactct gatcaaaatt ttgattacat gttcaaattg ctgatcatcg ggaacagcag 841 cgtcgggaaa acatccttcc tcttccgata cgctgatgat accttcacat ctgcttttgt 901 cagtacagtg ggcatcgact ttaaggtcaa aactgtctac aagaacgaca agaggatcaa 961 gcttcagatc tgggacacag ccggacagga acgatacagg actatcacca ctgcctacta 1021 cagaggggcc atgggcttca tcctaatgta tgacattact aacgaagagt cttttgctgc 1081 tgtacaggac tgggcgaccc agattaagac atactcgtgg gacaatgcac aggtgatcct 1141 cgctggaaac aaatgtgaca tggaggagga gaggatagtg tctgtggaca gcggcagact 1201 gctggcagaa cagctcggat ttgagttttt cgagaccagt gcgaaggaca acatcaacgt 1261 gaagcagact tttgaccgcc ttgtggatat aatctgcgac aagatgtccg acactctgga 1321 ggccgatcct gcggcggccg gcgggactcc cagcgccaga ctgaccgaga cccctgcacc 1381 ccctcagccg tcagactgca gctgctgatc ggctcaaccc aactttatta tacatagttg 1441 ataattcact ggccgtcgtt ttacggtacc atcgatgatg atccagacat gataagatac 1501 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa 1561 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac 1621 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc 1681 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctctagatca gatctgcgaa 1741 gatacggcca cgggtgctct tgatcctgtg gctgattttg gactgtgctg ctcgcagctg 1801 ctgatgaatc acatacttcc tccattttct tccactgatt gactgttata atttccctaa 1861 tttccaggtc aaggtgctgt gcattgtggt aatagatgtg acatgacgtc acttccaaag 1921 gaccaatgaa catgtctgac caatttcata taatgtgaaa acgattttca taggcagaat 1981 aaataacatt taaattaaac tgggcatcag cgcaattcaa ttggtttggt aatagcaagg 2041 gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa taaaattgta 2101 aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta aaagtattga 2161 tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag tacagtaatc 2221 aagtaaaatt actcaagtac tttacacctc tggttcttga ccccctacct tcagcaagcc 2281 cagcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc 2341 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa 2401 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 2461 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 2521 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 2581 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 2641 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 2701 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 2761 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 2821 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 2881 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 2941 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 3001 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 3061 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 3121 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 3181 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 3241 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 3301 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 3361 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 3421 cacgttaagg cattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 3481 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 3541 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 3601 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 3661 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 3721 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 3781 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 3841 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 3901 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 3961 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 4021 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 4081 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 4141 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 4201 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 4261 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 4321 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 4381 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 4441 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 4501 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt 4561 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 4621 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 4681 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 4741 cgatggccca cgctggctaa gaactcatca gcctccccgg tccatctacc cacgtaccaa 4801 tgcaccaatt ggccacaatg acggctacta catggtgcca ttccttcctc tttataggaa 4861 tggagactac ctcctgtcca acaatgctct tggatacgag tacgcctacc tgttggaccc 4921 aggtcattgc acaacaccag aaatgccctc tgatctgcaa aagacgtgaa tatctgttca 4981 gacacccata tccactctgt tccacacagg tcagaggttt gtccaggagt tcttgacaga 5041 ggtgtaaaaa gtactcaaaa attttactca agtgaaagta caagtactta gggaaaattt 5101 tactcaatta aaagtaaaag tatctggcta gaatcttact tgagtaaaag taaaaaagta 5161 ctccattaaa attgtacttg agtattaagg aagtaaaagt aaaagcaaga aagaaaacta 5221 gagattcttg tttaagcttt taatctcaaa aaacattaaa tgaaatgcat acaaggtttt 5281 atcctgcttt agaactgttt gtatttaatt atcaaactat aagacagaca atctaatgcc 5341 agtacacgct actcaaagtt gtaaaacctc agatttaact tcagtagaag ctgattctca 5401 aaattgttag tgtcaagcct agctcttttg gggctgaaaa gcaatcctgc agtgctgaaa 5461 agcctctcac aggcagccga tgcgggaaga ggtgtattag tcttgataga gaggctgcaa 5521 atagcaggaa acgtgagcag agactccctg gtgtctgaaa cacaggccag atgggccctc 5581 gagcaggaaa cagctatgac catgattacg ccaagctatc aactttgtat agaaaagttg 5641 gcacgcgcag attccgtgtg ggcctggtaa tatcttacca ttttaagcta ttttaagatt 5701 agggaaaaaa attaaataaa actactttaa aattactttc cttaaaaagc aacatattta 5761 tttagttttt gtttaactgg ctcaagattg taatatttta cttttacatg tttttttaag 5821 tacctaaatg tgttcaaaag taaagtatgt tattgttttt tttttttaat gttagctaag 5881 ctttaaattg taatatatta cttttaaaag tgacttttta aaaatgtaaa taaaaaagta 5941 actataaatg tgtaatgcat gttatttctg tgggttttta attttcatct caactgggct 6001 tacaatttta acataataaa aatcaaatca aatcaaatca ggcccttatt tgtcacatac 6061 actcttgtat actgaaataa actaaatgtt gagaaaaatt taaatacacc aaaataacgc 6121 ataccttttt tgtaattacc taagtaaaac atttagttcc ttttttaagt gactcaagat 6181 tgtaatctat taattgtaaa agataaaact agtgatcaat ctgacgtgct tctatgcagc 6241 ctgtgagatt aattgtgatg tttaattatt aaacaattta aatggatcaa acatttcaca 6301 tgtgcttatt gtgtccaaca aaaccagcag aagcacttaa aaatattgca tgacaacatg 6361 agtttttaat gtaaattata tatacttcct aaacgaagtt aatcatattt ctaaaataat 6421 aatgtcattg aaatatttaa ataaatgata agctttagtt gattcaacat aaaattgaaa 6481 gcaataagga cttttttttt ttacagcggt gaagcaaaag catgtaataa tcatataata 6541 taataaataa ataaataaat aatggctctt ggtgcaaata tgaacacttt aaataaatat 6601 agaataaata tgatcaataa gtcataacga tgtgttttat gttgattata atttaaacat 6661 gtttctaacg caagttttaa aaactgtttt aaagtattat tttaagttat aagatcaatt 6721 tgactcaaca ttttgtagtg aaaatatgtt gtgtaaaatt aatataatat acactataaa 6781 aatgcagggt tccacacaat acattaatgt tgtcccaaca caaatcgatt aagttaacaa 6841 atatatgtgg attgaacata aaaaattaag ttgtcccaat gaaatctcaa gaattgtgtt 6901 gtttcagctc attttaaata agtagtttta acaagtaaaa aataacatat ttttgagtgt 6961 gtatatatat atattacaaa atataataat ataaaatgta aaagatataa accatgctac 7021 atatatttaa gagctatttt gcacattact tttttgcaag ccattttagt gtttttttta 7081 caaatatata ttagctaaat atacaatatc ttaaatatac aattaatcag acatgcaatt 7141 aatgcattta acctacataa aaatacatga tcaacatatg ttttatgctg attacaacta 7201 aatcaaacgt aatgaaatta atttataaaa gcgatttaaa tatgtcaata ttttaagtta 7261 aaatcacttg aaacctacta taaatgaact taaattaaaa aaatctcctt taattccaat 7321 gcaacagtgt tgttttttca gctgaggacc cccggtgcag atggttggcc cccggcccgc 7381 tattctaagg aagcctttct taattttagg gcggcgctgc atgcgctcag atcagattca 7441 cgggtggaga gggttatttt ctctttttta ttttttcttg cctgctcata tttgggcctg 7501 tgtgaacccc aggctccgcc tcggccccca ggagccgctc ttatttaaag gggatcctcg 7561 ccttcagtct tcgtcccagt ggaagcatca gtcttgtgat aacaacttgg tgagtcctac 7621 agaccagcgc tcgactctag gctttaaatg cattgatttc tctggattag ctaccgggag 7681 agttttaact ttgcttatta gtcttgtgct cgaactgcta ggtttaaaga gtgttttctc 7741 taatgcatgc atgaagcgac agctgaaaac cagaaataga gcgtttgtcc tcctgttaaa 7801 ctctagcctt tctttggcat ttgtcagtcc gagtcttcag ctgccggcag gagattgggt 7861 ttcagtaaca cttgctgagc acaaactaca cgtgctttct tcagatttaa ttcattatta 7921 atcattctag atttaaatga tgttcttact cattgctgat tattttagcc ttgcggagca 7981 gattaaatga gcacgtgttt cattaagatt gatttaaata taattaatat ttcaccactg 8041 tagttttaaa attgccttct aagtcatggt tgatttttaa taggtatcgg ctctaattag 8101 attactattt tggtttttaa atgttatttc tatttcattt gtaggctcgt ttttaatgtg 8161 attttattag attctagttt taaaatcatc tcttagcctt aatgagcaga ctgaatgaac 8221 atgtttcatt aagattaact tcattataat gaattattta atcattctag tgggttataa 8281 atcagctcta tttaaattaa ttgtcgtgtt tatttctatt taatttttga ttattttttc 8341 agggtcgttt ttatgtgatt ttttcccgtt tctaatttta aactaattgt ttgattttta 8401 tatgattcga ctctttttaa atgaattgtt ttgctttttt tattgagcat atatatatta 8461 tacaatttca atcaacaatt gttgtgcttt ttaaatgttg tttctatttc atttctaggg 8521 tcgattttat tcgattctag ttttaaaatc gtgattttta acttctaacc ttactgagca 8581 gactgaatga acatgtttca tcaagattaa cttcattata gttaattatt gaatcgtgct 8641 agtggtttat aaatcagctc tatttaaatt aatcatcatc attatttcta tttaattttt 8701 attatttctt taggattatt tttatgtgat ttttttcacg ttctagtttt aaactcaatg 8761 ctcattactg ctcacccttg ctgagcagtc tttcgttaag attaaattaa ttaataacca 8821 ttctagtttt aaaacagtgt cttaaggcat agctgatatt ttacaagttt actatttaat 8881 atatctcctt tcttttttta gggtcgattt catctgattt gattaatttt tattatcttc 8941 actaagcttt ttgttctagg ttttattctc agtattatta ctgttatatt tgaactgaat 9001 ttctcctcat agtttattca ttttatttat ttattttttg tttattcgct attttgtatt 9061 ttagttttta attagaaatt ttattagaat cgttttgaac tagtttttat gcactctaac 9121 attaatatta aatcataact attattttta ttataattta tttgtgttta tctttttgtg 9181 tagctttcat gtgtttttat tttagtttta ttatgtttgc attgaatttt ttggactagt 9241 ttatataacg atatacagaa aagccatgac aacaaatgtt tttataatac ctattaagta 9301 taagctaatc agatttccac tattctgttc aaagttaaca ttattagcca ttctgatgga 9361 tcatttgaca tttgtcatat gtctttatga acactgctct ttcagatgcc acttttaaag 9421 ctgtttttta cagtaaatga acgtttatta ttatgacatt atttcctaag gcctctagag 9481 tacagcagga tgtggcgtgt gcattaccgt gtaattatgc tgactgtaaa tgtgcattta 9541 tagactgtca gcatgtctga ctgtgcgctg ttgtgattgg tcctgcacag accaag //