LOCUS Exported 9716 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab34b. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab34b SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9716) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9716) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9716 /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(6613..6633) /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(6641..6657) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 6673..6807 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6697..6716) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6751..6770 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(7479..7495) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7479..7495) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(7532..7552) /label=T3 /note="T3 promoter, forward primer" promoter complement(7532..7550) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7571..7587) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7584..7606) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7595..7611 /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(7619..7649) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7664..7685 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7802..7819) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7973..8561) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8053..8072) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8732..9592) /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 9355..9374 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9593..9697) /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 tggtgatgaa ttctttaccc cctgtgaaag gagaccggat 5881 catctgtgaa ctccctttgt attttacaag agatgctgca gtgcacacca aagatggctt 5941 caatattact gttagagagg cttgtcaagg gcagaaaaca ggattaaatg tagccaaagt 6001 cattgtcgtt ggtgatgttg gggtcgggaa gacctgttta ataaacagat tctgcaaaga 6061 ctcatttgac agaacgtaca aggcaactat tggagtggat tttgagatgg agagatttga 6121 agttctcggg gtacccttca gtttgcagct gtgggacaca gcaggtcagg aaagatttcg 6181 atgcattgcc tccacatact acagaggagc gcaagccatc atcgttgtgt ttgacctgag 6241 caactatgag tctctggatc acgcgagaga atggcttgaa gatgccatga gggacaatga 6301 cccgtccagt gtcctgctct tcctcgtggg aaccaagaag gatcttagtc atccagatct 6361 gttggcccaa atggagcagg aggccattcg attgtctgag gagatcagag cagagtactg 6421 ggcagtttct gctttgtcag gggcgagtgt gagagatttc ttcctgcggg ttgcttcgct 6481 tacatttgag gccactgttt tatctgagct ggagaagaac aattctagac gagccggtga 6541 tatcgtcaaa atctccaata attgtgacaa caattttaag aagaggcaat ccagctgctg 6601 ccattgactg aacaacttta ttatacatag ttgataattc actggccgtc gttttacggt 6661 accatcgatg atgatccaga catgataaga tacattgatg agtttggaca aaccacaact 6721 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta 6781 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag 6841 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg 6901 gctgattatg atcctctaga tcagatctgc gaagatacgg ccacgggtgc tcttgatcct 6961 gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact tcctccattt 7021 tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc tgtgcattgt 7081 ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct gaccaatttc 7141 atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta aactgggcat 7201 cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg atctccaaaa 7261 aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa 7321 aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa 7381 aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag tactttacac 7441 ctctggttct tgacccccta ccttcagcaa gcccagcaga tccactagtt ctagagcggc 7501 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg 7561 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 7621 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 7681 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 7741 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 7801 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 7861 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 7921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 7981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 8041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 8101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 8161 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 8221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 8281 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 8341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 8401 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 8461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 8521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 8581 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 8641 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 8701 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 8761 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 8821 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 8881 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 8941 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 9001 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 9061 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 9121 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 9181 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 9241 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 9301 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 9361 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 9421 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 9481 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 9541 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 9601 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 9661 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtg //