LOCUS Exported 9226 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION heat shock inducible transgenesis. ACCESSION . VERSION . KEYWORDS hsp70:EB3-mKate2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9226) AUTHORS Icha J, Kunath C, Rocha-Martins M, Norden C TITLE Independent modes of ganglion cell translocation ensure correct lamination of the zebrafish retina. JOURNAL J Cell Biol. 2016 Oct 24;215(2):259-275. PUBMED 27810916 REFERENCE 2 (bases 1 to 9226) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9226 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(7..462) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(144..165) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 356..375 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 589..611 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 603..620 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 604..620 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 630..649 /label=T7 /note="T7 promoter, forward primer" promoter 630..648 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1324..1344 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" protein_bind 2864..2888 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3752..4450 /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(4452..4476) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" polyA_signal 4481..4615 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4505..4524) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4559..4578 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(6989..7005) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6989..7005) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(7042..7062) /label=T3 /note="T3 promoter, forward primer" promoter complement(7042..7060) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7081..7097) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7081..7097) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7094..7116) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7105..7121 /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(7129..7159) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7174..7195 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7312..7329) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7483..8071) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7563..7582) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8242..9102) /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 8865..8884 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9103..9207) /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 tggcccacta cgtgaaccat 241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc 481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga 541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac 601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta 661 caggtcattg cacaacacca gaaatgccct ctgatctgca aaagacgtga atatctgttc 721 agacacccat atccactctg ttccacacag gtcagaggtt tgtccaggag ttcttgacag 781 aggtgtaaaa agtactcaaa aattttactc aagtgaaagt acaagtactt agggaaaatt 841 ttactcaatt aaaagtaaaa gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt 901 actccattaa aattgtactt gagtattaag gaagtaaaag taaaagcaag aaagaaaact 961 agagattctt gtttaagctt ttaatctcaa aaaacattaa atgaaatgca tacaaggttt 1021 tatcctgctt tagaactgtt tgtatttaat tatcaaacta taagacagac aatctaatgc 1081 cagtacacgc tactcaaagt tgtaaaacct cagatttaac ttcagtagaa gctgattctc 1141 aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa agcaatcctg cagtgctgaa 1201 aagcctctca caggcagccg atgcgggaag aggtgtatta gtcttgatag agaggctgca 1261 aatagcagga aacgtgagca gagactccct ggtgtctgaa acacaggcca gatgggccga 1321 tatcaacttt gtatagaaaa gttgcttcag gggtgtcgct tggttatttc caaaaatcaa 1381 attaatttta ttaaactatt agaacgagca tgttttgtct atatgctaca gaagataaaa 1441 aataatagga gttaacagtt ataaaacaac acactttgtt tctattgatt gttgaccaca 1501 ctggggtctc attaagttag attaaagaca cactaactgg gtcaaaagca gcagattgat 1561 ttcatagcac cagggtaaac tttctaacac ttttacggca atcatataca ttaaaattaa 1621 atacagacca cgactgaaca aggaggatga tctccaatat taaacaaaga gacttgtgcc 1681 tatttctctg agggtaaaca tgacctctca agttagcaag ttggttttaa cactacaaaa 1741 atagttaaga ctgcaatccc agaataaagt attggtttta accaatcaat atagtacagt 1801 aaacatccat ttgttttgtt gaaacgttaa acaaatctga ccaaagctat tagcttatat 1861 aaaacaggtt tgccttctat gtagctgaaa acaccacagg cccgattttg ctactgtgta 1921 aaacatttca gcaagatttt tttattgcat ttttttttac tgaatcgttc aaacatttta 1981 tcattttagt ttgttcattc attgcaactg gaaaaacaac acatcacaca accgcacata 2041 tttcagcaat aagtacaata aaacactcaa ataaaaaaaa cattttaaat ctctttgtat 2101 ttttgaccgc tgtttcgcgt aatttcacgg taaaactctg gaaatctcca ctacattcct 2161 ctcagcggct cctctcaatg acagctgaag aagtgacgcg gctgcctgct gtgttttgat 2221 tggtcgaatt cactggaggc ttccagaaca gtgtagagtc tgaacgggtg cgcgctctgc 2281 tgtatttaaa gggcgaaaga gagaccgcag agaaactcaa ccgaagagaa gcgacttgac 2341 aaagaagaaa agagcagcct gacaggactt tttccccgac gaggtgttta ttcgctctat 2401 ttaagaatct actgtaaggt aagtctcaat atattgtact ctattggcta atcagaatta 2461 tatagagatt atatgtactt aatgtcaaaa aatcaacttt gtatatgtaa tctttttaca 2521 tgtggactgc ctatgttcat cttattttag gtctactaga aaattatatt tcccgttttc 2581 acaataagga tttttaaaaa aagcaatgaa cagacgggca tttactttat gttgctgaca 2641 ttattttata tgagcataat aaccataaat actagcaaat gtcctaaatg aatttgtgtt 2701 aatgttgtct acaaaagaaa attagcgttt tacttgtaca actaataata acttggttat 2761 taagagaatt tcacttgttg actagaaaaa tcctttcata atgaaacaat tgcaccataa 2821 attgtataaa tataaaatta attctaattg tttttttttc ctgccaagtt tgtacaaaaa 2881 agcaggctgg atggccgtca atgtgtactc cacatctgtg accagtgaaa atctgagtcg 2941 ccatgatatg cttgcatggg tcaacgactc cctgcacctc aactatacca agatagaaca 3001 gctttgttca ggggcagcct actgccagtt catggacatg ctcttccccg gctgtgtgca 3061 cttgaggaaa gtgaagttcc aggccaaact agagcatgaa tacatccaca acttcaaggt 3121 gctgcaagca gctttcaaga agatgggtgt tgacaaaatc attcctgtag agaaattagt 3181 gaaaggaaaa ttccaagata attttgagtt tattcagtgg tttaagaaat tctttgacgc 3241 aaactatgat ggaaaggatt acaaccctct gctggcgcgg cagggccagg acgtagcgcc 3301 acctcctaac ccaggtgatc agatcttcaa caaatccaag aaactcattg gcacagcagt 3361 tccacagagg acgtccccca caggcccaaa aaacatgcag acctctggcc ggctgagcaa 3421 tgtggccccc ccctgcattc tccggaagaa tcctccatca gcccgaaatg gcggccatga 3481 gactgatgcc caaattcttg aactcaacca acagctggtg gacttgaagc tgacagtgga 3541 tgggctggag aaggaacgtg acttctactt cagcaaactt cgtgacatcg agctcatctg 3601 ccaggagcat gaaagtgaaa acagccctgt tatctcaggc atcattggca tcctctatgc 3661 cacagaggaa ggattcgcac cccctgagga cgatgagatt gaagagcatc aacaagaaga 3721 ccaggacgag tacaccggtt catcgggatc catggtgagc gagctgatta aggagaacat 3781 gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga 3841 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg 3901 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt 3961 catcaaccac acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac 4021 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag 4081 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa 4141 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc 4201 cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca 4261 cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat 4321 gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac 4381 ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg 4441 gcacagatga gacccagctt tcttgtacaa agttgatgta gatccagaca tgataagata 4501 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga 4561 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa 4621 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt 4681 cgcggccgcg gcgccaatgc attgggccct cgagcatatg agatctgtcc cgatctttct 4741 cttctgtgct gtcaatttag agtagttttt gaggtaattt gggtgcattc tctgtaaaaa 4801 caacaaataa atatttatat tgtatgcaat aaatgtaaca attacacata cactctctga 4861 ttacagcatt aaagggttag ttcacccaaa aatgaaaaga tgtcattaat gactcaccct 4921 catgtcgttc caagcccgta acacctccgt tcatcttcgg aacacagttt aagatatttt 4981 agatttagtc cgagagcttt ctgtccctcc actgaaaatg tatgtacggt atactgtcca 5041 tgtccagaaa ggtaataaaa acatcaaagt agtccatgtg acatcagtgg gttaggtaga 5101 attatttgaa gcatcgaaaa tacattttgg tccaaaaata acaaaaccta cgactttatt 5161 cagcattgta gcgtcactgc gaagtcgtga acgcggattg acaacagacc cggaagagaa 5221 tacaatgccg aataaagtcg taggttttgt tatttttgga ccaaaatgta ttcgatgctt 5281 caaaaaattc taactaaccc actgatgtca catggactac tttgatgttt ttattacctt 5341 tctggacatg gacagtatac cgtacataca ttctcaatgg aggcacagaa agctctcgga 5401 ctaaatctaa aatatcttaa actgtgttct gaagatgaac ggaggtctta cgggcttgga 5461 acgacatgag ggcgagtcat taatgacatt attttcattt ttgggtgaac taacccttta 5521 ataaaactga catttgcagc caattgcaca ctttataatg ttaaatactt gttgatttta 5581 gaaatacagt agcatttagt gacaatcaaa cattaatttt atttctgatt atattgacaa 5641 aaacacagga ctgtaaggct gacatgacag ggtgaattga tttcattaca ttagttttga 5701 tggtaacact tttacaataa ggttcgttag ttaactacat tagttaacat gaactaataa 5761 tgaactgcac gtatacatgt taatttcaac atttactaat actttatatc ttgttaacat 5821 tagttaatgc actgtgaact aacatgaaca aacaatgaac atctgtattt ctattaacta 5881 acgttaacaa agattaacag atacagtaac aaatgtattg ctcatggtta tttaatgtta 5941 gttaatacat gaactaatca acctcattgt aaagtgttac cgttttgaca atacaacatg 6001 tttggaaaaa aatttctcgc ttggttttct tgtgttttgc atgaattcaa acacaatgag 6061 agcacggaat gataatgatt attcaaagct gcaataggcg ccaaatttcc cgtggaccta 6121 acgttaccaa ttacaaatgc attttagcat tttattacgt gagccttata ttgtcaatcg 6181 catgcattag tgaaaatagg cctactacac atataaaagg cgcgccatca acgtcaaaac 6241 atgcacaccc aaaaaaaaaa aaaaaaaaaa aaaaaaaaac tatcagtaaa acaaaatact 6301 taatgtactt acctcaatat gcttccttag gtttgatggc gaacttttga aggccgatat 6361 ttctttatta agcgggagac agaggacaca tttcatcttg aatgaatctt tatttacacc 6421 acttaaagaa aagaattcgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt 6481 ttggactgtg ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg 6541 attgactgtt ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat 6601 gtgacatgac gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg 6661 aaaacgattt tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt 6721 caattggttt ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact 6781 ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa 6841 ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa 6901 aataatactt aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct 6961 tgacccccta ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg 7021 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg 7081 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7141 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7201 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7261 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 8041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 8101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 9001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 9061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtg //