LOCUS Exported 9581 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Muscle specific zebrafish expression of mKate2 fused to rab39ba. ACCESSION . VERSION . KEYWORDS actc1b-mKate2-rab39bb SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9581) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 9581) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9581 /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(1405..1425) /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(1433..1449) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 1465..1599 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1489..1508) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1543..1562 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2271..2287) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2271..2287) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2324..2344) /label=T3 /note="T3 promoter, forward primer" promoter complement(2324..2342) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2363..2379) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2376..2398) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2387..2403 /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(2411..2441) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2456..2477 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2594..2611) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2765..3353) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2845..2864) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3524..4384) /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 4147..4166 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4385..4489) /gene="bla" /label=AmpR promoter primer_bind complement(4652..4673) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5569..5585 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5605..5625 /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 ggtggaggcg atttggcttt accagtttcg 781 gctgattgtg attggggact caactgttgg aaagtcgtgt ttaattaggc gatttacaga 841 aggccgtttc gcgcaggtgt cggacccgac agtcggggtc gattttttct cccgcctggt 901 ggagattgaa cccggaaaac gcattaaact gcagatctgg gacactgcag gccaggaacg 961 tttcaggtct attactcgag catactaccg taactctgtg ggcggcctgc tgctcttcga 1021 catcaccaac cgccgctctt tccagaacgt tcacgagtgg ctagaggagg cgcgcagtca 1081 cgtgcagccc cacagcatcg tcttcctgct agtcgggcac aaatgtgatc tggaaccaca 1141 gcgccaggtg agccagcaag aggccgagaa gcttgcagcc gcatacggta tgcgctacgt 1201 ggagacttcg gcacgtgacg ccataaacgt ggagagggct tttacagagc taacgcggga 1261 tatatttgag ctggtcaaga atggggagat tagcatccag gaaggctggg agggagtgaa 1321 gagcgggttc gtaccaaacg tggtgcattc gtcagaagaa gtgacaaagg gtgatcgccg 1381 gtgcttctgt tgaatccgca atcacaactt tattatacat agttgataat tcactggccg 1441 tcgttttacg gtaccatcga tgatgatcca gacatgataa gatacattga tgagtttgga 1501 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt 1561 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat 1621 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac 1681 aaatgtggta tggctgatta tgatcctcta gatcagatct gcgaagatac ggccacgggt 1741 gctcttgatc ctgtggctga ttttggactg tgctgctcgc agctgctgat gaatcacata 1801 cttcctccat tttcttccac tgattgactg ttataatttc cctaatttcc aggtcaaggt 1861 gctgtgcatt gtggtaatag atgtgacatg acgtcacttc caaaggacca atgaacatgt 1921 ctgaccaatt tcatataatg tgaaaacgat tttcataggc agaataaata acatttaaat 1981 taaactgggc atcagcgcaa ttcaattggt ttggtaatag caagggaaaa tagaatgaag 2041 tgatctccaa aaaataagta ctttttgact gtaaataaaa ttgtaaggag taaaaagtac 2101 ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt attgattttt aattgtactc 2161 aagtaaagta aaaatcccca aaaataatac ttaagtacag taatcaagta aaattactca 2221 agtactttac acctctggtt cttgaccccc taccttcagc aagcccagca gatccactag 2281 ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa 2341 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 2401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 2461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 2521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 2581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 2641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 2701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 2761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 2821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 2881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 2941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 3001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 3061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 3121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 3181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 3241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 3301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 3361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taaggcattt 3421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 3481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 3541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 3601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 3661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 3721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 3781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 3841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 3901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 3961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 4021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 4081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 4141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 4201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 4261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 4321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 4381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 4441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 4501 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 4561 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 4621 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 4681 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccacgctg 4741 gctaagaact catcagcctc cccggtccat ctacccacgt accaatgcac caattggcca 4801 caatgacggc tactacatgg tgccattcct tcctctttat aggaatggag actacctcct 4861 gtccaacaat gctcttggat acgagtacgc ctacctgttg gacccaggtc attgcacaac 4921 accagaaatg ccctctgatc tgcaaaagac gtgaatatct gttcagacac ccatatccac 4981 tctgttccac acaggtcaga ggtttgtcca ggagttcttg acagaggtgt aaaaagtact 5041 caaaaatttt actcaagtga aagtacaagt acttagggaa aattttactc aattaaaagt 5101 aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa aagtactcca ttaaaattgt 5161 acttgagtat taaggaagta aaagtaaaag caagaaagaa aactagagat tcttgtttaa 5221 gcttttaatc tcaaaaaaca ttaaatgaaa tgcatacaag gttttatcct gctttagaac 5281 tgtttgtatt taattatcaa actataagac agacaatcta atgccagtac acgctactca 5341 aagttgtaaa acctcagatt taacttcagt agaagctgat tctcaaaatt gttagtgtca 5401 agcctagctc ttttggggct gaaaagcaat cctgcagtgc tgaaaagcct ctcacaggca 5461 gccgatgcgg gaagaggtgt attagtcttg atagagaggc tgcaaatagc aggaaacgtg 5521 agcagagact ccctggtgtc tgaaacacag gccagatggg ccctcgagca ggaaacagct 5581 atgaccatga ttacgccaag ctatcaactt tgtatagaaa agttggcacg cgcagattcc 5641 gtgtgggcct ggtaatatct taccatttta agctatttta agattaggga aaaaaattaa 5701 ataaaactac tttaaaatta ctttccttaa aaagcaacat atttatttag tttttgttta 5761 actggctcaa gattgtaata ttttactttt acatgttttt ttaagtacct aaatgtgttc 5821 aaaagtaaag tatgttattg tttttttttt ttaatgttag ctaagcttta aattgtaata 5881 tattactttt aaaagtgact ttttaaaaat gtaaataaaa aagtaactat aaatgtgtaa 5941 tgcatgttat ttctgtgggt ttttaatttt catctcaact gggcttacaa ttttaacata 6001 ataaaaatca aatcaaatca aatcaggccc ttatttgtca catacactct tgtatactga 6061 aataaactaa atgttgagaa aaatttaaat acaccaaaat aacgcatacc ttttttgtaa 6121 ttacctaagt aaaacattta gttccttttt taagtgactc aagattgtaa tctattaatt 6181 gtaaaagata aaactagtga tcaatctgac gtgcttctat gcagcctgtg agattaattg 6241 tgatgtttaa ttattaaaca atttaaatgg atcaaacatt tcacatgtgc ttattgtgtc 6301 caacaaaacc agcagaagca cttaaaaata ttgcatgaca acatgagttt ttaatgtaaa 6361 ttatatatac ttcctaaacg aagttaatca tatttctaaa ataataatgt cattgaaata 6421 tttaaataaa tgataagctt tagttgattc aacataaaat tgaaagcaat aaggactttt 6481 tttttttaca gcggtgaagc aaaagcatgt aataatcata taatataata aataaataaa 6541 taaataatgg ctcttggtgc aaatatgaac actttaaata aatatagaat aaatatgatc 6601 aataagtcat aacgatgtgt tttatgttga ttataattta aacatgtttc taacgcaagt 6661 tttaaaaact gttttaaagt attattttaa gttataagat caatttgact caacattttg 6721 tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc agggttccac 6781 acaatacatt aatgttgtcc caacacaaat cgattaagtt aacaaatata tgtggattga 6841 acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc agctcatttt 6901 aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata tatatatatt 6961 acaaaatata ataatataaa atgtaaaaga tataaaccat gctacatata tttaagagct 7021 attttgcaca ttactttttt gcaagccatt ttagtgtttt ttttacaaat atatattagc 7081 taaatataca atatcttaaa tatacaatta atcagacatg caattaatgc atttaaccta 7141 cataaaaata catgatcaac atatgtttta tgctgattac aactaaatca aacgtaatga 7201 aattaattta taaaagcgat ttaaatatgt caatatttta agttaaaatc acttgaaacc 7261 tactataaat gaacttaaat taaaaaaatc tcctttaatt ccaatgcaac agtgttgttt 7321 tttcagctga ggacccccgg tgcagatggt tggcccccgg cccgctattc taaggaagcc 7381 tttcttaatt ttagggcggc gctgcatgcg ctcagatcag attcacgggt ggagagggtt 7441 attttctctt ttttattttt tcttgcctgc tcatatttgg gcctgtgtga accccaggct 7501 ccgcctcggc ccccaggagc cgctcttatt taaaggggat cctcgccttc agtcttcgtc 7561 ccagtggaag catcagtctt gtgataacaa cttggtgagt cctacagacc agcgctcgac 7621 tctaggcttt aaatgcattg atttctctgg attagctacc gggagagttt taactttgct 7681 tattagtctt gtgctcgaac tgctaggttt aaagagtgtt ttctctaatg catgcatgaa 7741 gcgacagctg aaaaccagaa atagagcgtt tgtcctcctg ttaaactcta gcctttcttt 7801 ggcatttgtc agtccgagtc ttcagctgcc ggcaggagat tgggtttcag taacacttgc 7861 tgagcacaaa ctacacgtgc tttcttcaga tttaattcat tattaatcat tctagattta 7921 aatgatgttc ttactcattg ctgattattt tagccttgcg gagcagatta aatgagcacg 7981 tgtttcatta agattgattt aaatataatt aatatttcac cactgtagtt ttaaaattgc 8041 cttctaagtc atggttgatt tttaataggt atcggctcta attagattac tattttggtt 8101 tttaaatgtt atttctattt catttgtagg ctcgttttta atgtgatttt attagattct 8161 agttttaaaa tcatctctta gccttaatga gcagactgaa tgaacatgtt tcattaagat 8221 taacttcatt ataatgaatt atttaatcat tctagtgggt tataaatcag ctctatttaa 8281 attaattgtc gtgtttattt ctatttaatt tttgattatt ttttcagggt cgtttttatg 8341 tgattttttc ccgtttctaa ttttaaacta attgtttgat ttttatatga ttcgactctt 8401 tttaaatgaa ttgttttgct ttttttattg agcatatata tattatacaa tttcaatcaa 8461 caattgttgt gctttttaaa tgttgtttct atttcatttc tagggtcgat tttattcgat 8521 tctagtttta aaatcgtgat ttttaacttc taaccttact gagcagactg aatgaacatg 8581 tttcatcaag attaacttca ttatagttaa ttattgaatc gtgctagtgg tttataaatc 8641 agctctattt aaattaatca tcatcattat ttctatttaa tttttattat ttctttagga 8701 ttatttttat gtgatttttt tcacgttcta gttttaaact caatgctcat tactgctcac 8761 ccttgctgag cagtctttcg ttaagattaa attaattaat aaccattcta gttttaaaac 8821 agtgtcttaa ggcatagctg atattttaca agtttactat ttaatatatc tcctttcttt 8881 ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa gctttttgtt 8941 ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc ctcatagttt 9001 attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt ttttaattag 9061 aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa tattaaatca 9121 taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct ttcatgtgtt 9181 tttattttag ttttattatg tttgcattga attttttgga ctagtttata taacgatata 9241 cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc taatcagatt 9301 tccactattc tgttcaaagt taacattatt agccattctg atggatcatt tgacatttgt 9361 catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt ttttacagta 9421 aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag caggatgtgg 9481 cgtgtgcatt accgtgtaat tatgctgact gtaaatgtgc atttatagac tgtcagcatg 9541 tctgactgtg cgctgttgtg attggtcctg cacagaccaa g //