LOCUS Exported 7349 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION phosphatase-dead control for 5HT6-YFP-Inpp5e, CAAX-null. ACCESSION . VERSION . KEYWORDS 5HT6-YFP-Inpp5e(D477N) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7349) AUTHORS Phua SC, Chiba S, Suzuki M, Su E, Roberson EC, Pusapati GV, Setou M, Rohatgi R, Reiter JF, Ikegami K, Inoue T TITLE Dynamic Remodeling of Membrane Composition Drives Cell Cycle through Primary Cilia Excision. JOURNAL Cell. 2017 Jan 12;168(1-2):264-279.e15. doi: 10.1016/j.cell.2016.12.032. Epub 2017 Jan 12. PUBMED 28086093 REFERENCE 2 (bases 1 to 7349) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7349 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 6..594 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 495..514 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 769..1072 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1073..1276 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1227..1247 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1305..1314 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 2661..2670 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2667..3383 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2712..2733) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2973..2992) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3320..3341 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4845..4966 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4882..4901) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4936..4955 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4973..5428) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5110..5131) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5322..5341 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5455..5559 /gene="bla" /label=AmpR promoter promoter 5561..5918 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5584..5604) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5769..5904 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5831..5850 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5953..6747 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6007..6026) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6617..6636 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6935..6954) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6979..7026 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 61 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 181 cacttcaaga actctgtatc accgcctaca tacctcgctc tgctaatcct gttaccagtg 241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 661 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata 721 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 781 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 841 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 901 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 961 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 1021 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 1081 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 1141 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 1201 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 1261 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgccacc atggttccag 1321 agcccggccc tgtcaacagt agcaccccag cctggggccc cgggccaccg ccagccccag 1381 ggggcagcgg ctgggtggcc gctgcgctgt gcgtggtcat cgtgctgacg gcagccgcca 1441 attcgctgct gatcgcgctc atctgcacgc agcccgcgct gcgcaacacg tctaacttct 1501 tcctggtgtc gctcttcacg tcggacctga tggtgggatt ggtggtgatg cccccagcca 1561 tgctgaacgc gctgtatggg cgctgggtac tagctcgcgg cctctgtctg ctttggaccg 1621 ccttcgacgt gatgtgctgc agcgcctcca ttctcaacct ctgcctcatc agcctggacc 1681 gctacctgct catcctctcg ccgctgcgct acaagctgcg catgacagcc ccgcgtgccc 1741 tggcgctcat cctgggtgcc tggagcctcg ccgcgctggc ctccttcctg cccctcttgc 1801 tgggctggca tgaactgggc aaagctcgaa catctgcccc gggccagtgc cgcctattgg 1861 ccagcctgcc ttatgtcctc gtggcgtccg gcgtcacctt tttcctgcct tcgggtgcca 1921 tctgcttcac ctactgcagg atcttgctgg ctgcccgcaa gcaggcggtg caagtggcct 1981 cgctcaccac gggcacggct acggctggcc aggccttgga aaccttgcag gtgcccagga 2041 caccacgccc agggatggag tctgctgaca gtaggcgtct gaccaccaag catagcagga 2101 aggccttgaa ggccagcctg accctcggca tcctgctgag catgttcttt gtcacctggc 2161 tgcccttctt tgtggccagc atagctcagg ccgtatgtga ctgcatctct ccaggtctct 2221 tcgatgtcct cacatggctg ggatactgta atagcaccat gaaccctatc atctatcccc 2281 tcttcatgcg ggacttcaag agggccctgg gcaggttcgt gccgtgtgtc cactgtcccc 2341 cggagcaccg ggccagcccc gcctccccct ccatgtggac ctctcacagt ggtgccaggc 2401 caggcctcag cctgcagcag gtgctgcccc tgcctctgcc acccaactca gattcagact 2461 cagcttcagg gggcacctcg ggcctgcagc tcacagccca gcttttgctg cctggagagg 2521 cgacccggga ccccccgcca cccaccaggg cccctactgt ggtcaacttc ttcgtcacag 2581 actctgtgga gcctgagata cggcagcatc cacttggttc ccccatgaac agtgctggtg 2641 gtagcgcagg aggaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg 2701 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 2761 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 2821 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct 2881 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 2941 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 3001 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 3061 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 3121 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 3181 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 3241 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc 3301 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 3361 tcggcatgga cgagctgtac aagtccggac tcagatctcg agctcaagct tcgaattcgt 3421 cggatcttgc agactacaag ctccgcgcgc agccgctcct ggtgcgggcc cacagcagcc 3481 tgggccccgg ccggccgcgg agccccctgg cctgcgacga ctgttccctt cgctcggcca 3541 aatcctcctt cagcctcctg gcgcccatcc gcagcaagga cgtccgcagc aggagttacc 3601 tggagggcag cctcctggcc agcggggccc tgttgggggc ggatgagctg gcccgctact 3661 tcccagaccg gaacgtggca ctcttcgtgg ccacctggaa catgcagggc cagaaggagc 3721 tcccgcccag cctggacgag ttcctgctcc cagccgaggc cgactatgcc caggacctgt 3781 atgtcatcgg ggtccaggag ggctgttctg acaggcggga gtgggagact cgtctgcagg 3841 agacgctggg cccgcactat gtgctgctgt cctcggcggc ccacggcgtg ctctacatgt 3901 cgctcttcat ccgcagggac ctcatctggt tctgctcaga ggtggagtgc tccacggtga 3961 ccacacgcat cgtgtctcag atcaagacca agggggcctt gggcatcagc ttcacctttt 4021 ttggcacttc cttcctcttc atcacgtccc acttcacctc aggtgacggg aaggtggcgg 4081 agcggctgct ggactacacc aggactgtac aagccctggt cctgcccaga aatgtgcccg 4141 acaccaaccc ctatcgctcc agcgcagcgg acgtcaccac ccgcttcgat gaggtgttct 4201 ggtttggaaa cttcaacttc cgcctgagtg gcgggcgcac agtcgtggac gccctcctgt 4261 gccagggcct ggtggtggac gtgccggcgc tgctgcagca cgaccagctc atccgggaga 4321 tgcggaaagg gtccatcttc aagggcttcc aggagccgga catccacttc ctcccatcat 4381 acaagtttga catcgggaag gacacgtacg acagcacctc caagcagagg acgccctcat 4441 acacggaccg cgtcttgtac agaagccgcc acaagggtga catctgtcct gtgagctact 4501 cttcctgccc cgggatcaag acgtccgacc accgccctgt gtatggcctc ttccgggtga 4561 aagtgaggcc ggggcgagac aacattccgt tggcagctgg caaatttgat agagaactgt 4621 acttactagg aattaaaaga cggatttcga aggagattca gaggcagcaa gcactacaga 4681 gtcagaactc cagcaccatc gcctccgttt cttgaggatc caccggatct agataactga 4741 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc 4801 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag 4861 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4921 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta 4981 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac 5041 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg 5101 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa 5161 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt 5221 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt 5281 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga 5341 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc 5401 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga 5461 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 5521 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc 5581 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 5641 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 5701 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 5761 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 5821 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt 5881 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga 5941 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 6001 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 6061 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 6121 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 6181 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 6241 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 6301 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 6361 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 6421 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 6481 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 6541 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 6601 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 6661 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 6721 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 6781 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 6841 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 6901 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg 6961 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt 7021 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac 7081 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg 7141 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt 7201 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 7261 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 7321 cgtcagaccc cgtagaaaag atcaaagga //