LOCUS Exported 8278 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression vector, containing a GFP tagged phosphatase dead version of INPP5D (D673G). ACCESSION . VERSION . KEYWORDS INPP5D-eGFP D673G mutant SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8278) AUTHORS Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA, McGarry L, Shanks E, Mostov KE, Bryant DM TITLE The phospholipid PI(3,4)P2 is an apical identity determinant. JOURNAL Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8. PUBMED 30487552 REFERENCE 2 (bases 1 to 8278) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8278 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 143..446 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 447..650 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 601..621 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 679..688 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" RBS 2278..2286 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" regulatory 4300..4309 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4306..5025 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4306..5022 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4351..4372) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4612..4631) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4959..4980 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 5148..5269 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5185..5204) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5239..5258 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5276..5731) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5413..5434) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5625..5644 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5758..5862 /gene="bla" /label=AmpR promoter promoter 5864..6221 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5887..5907) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6072..6207 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6134..6153 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6256..7050 /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(6310..6329) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6920..6939 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7238..7257) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7282..7329 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7658..8246 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8147..8166 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 61 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc 121 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 181 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 241 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 301 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 361 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 421 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 481 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 541 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 601 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga 661 accgtcagat ccgctagcgc caccatggtc ccctgctgga accatggcaa catcacccgc 721 tccaaggcgg aggagctgct ttccaggaca ggcaaggacg ggagcttcct cgtgcgtgcc 781 agcgagtcca tctcccgggc atacgcgctc tgcgtgctgt atcggaattg cgtttacact 841 tacagaattc tgcccaatga agatgataaa ttcactgttc aggcatccga aggcgtctcc 901 atgaggttct tcaccaagct ggaccagctc atcgagtttt acaagaagga aaacatgggg 961 ctggtgaccc atctgcaata ccctgtgccg ctggaggaag aggacacagg cgacgaccct 1021 gaggaggaca cagaaagtgt cgtgtctcca cccgagctgc ccccaagaaa catcccgctg 1081 actgccagct cctgtgaggc caaggaggtt cctttttcaa acgagaatcc ccgagcgacc 1141 gagaccagcc ggccgagcct ctccgagaca ttgttccagc gactgcaaag catggacacc 1201 agtgggcttc cagaagagca tcttaaggcc atccaagatt atttaagcac tcagctcgcc 1261 caggactctg aatttgtgaa gacagggtcc agcagtcttc ctcacctgaa gaaactgacc 1321 acactgctct gcaaggagct ctatggagaa gtcatccgga ccctcccatc cctggagtct 1381 ctgcagaggt tatttgacca gcagctctcc ccgggcctcc gtccacgtcc tcaggttcct 1441 ggtgaggcca atcccatcaa catggtgtcc aagctcagcc aactgacaag cctgttgtcg 1501 tccattgaag acaaggtcaa ggccttgctg cacgagggtc ctgagtctcc gcaccggccc 1561 tcccttatcc ctccagtcac ctttgaggtg aaggcagagt ctctggggat tcctcagaaa 1621 atgcagctca aagtcgacgt tgagtctggg aaactgatca ttaagaagtc caaggatggt 1681 tctgaggaca agttctacag ccacaagaaa atcctgcagc tgattaagtc acagaaattt 1741 ctgaataagt tggtgatctt ggtggaaaca gagaaggaga agatcctgcg gaaggaatat 1801 gtttttgctg actccaaaaa gagagaaggc ttctgccagc tcctgcagca gatgaagaac 1861 aagcactcag agcagccgga gcccgacatg atcaccatct tcatcggcac ctggaacatg 1921 ggtaacgccc cccctcccaa gaagatcacg tcctggtttc tctccaaggg gcagggaaag 1981 acgcgggacg actctgcgga ctacatcccc catgacattt acgtgatcgg cacccaagag 2041 gaccccctga gtgagaagga gtggctggag atcctcaaac actccctgca agaaatcacc 2101 agtgtgactt ttaaaacagt cgccatccac acgctctgga acatccgcat cgtggtgctg 2161 gccaagcctg agcacgagaa ccggatcagc cacatctgta ctgacaacgt gaagacaggc 2221 attgcaaaca cactggggaa caagggagcc gtgggggtgt cgttcatgtt caatggaacc 2281 tccttagggt tcgtcaacag ccacttgact tcaggaagtg aaaagaaact caggcgaaac 2341 caaaactata tgaacattct ccggttcctg gccctgggcg acaagaagct gagtcccttt 2401 aacatcactc accgcttcac gcacctcttc tggtttgggg atcttaacta ccgtgtggat 2461 ctgcctacct gggaggcaga aaccatcatc cagaaaatca agcagcagca gtacgcagac 2521 ctcctgtccc acgaccagct gctcacagag aggagggagc agaaggtctt cctacacttc 2581 gaggaggaag aaatcacgtt tgccccaacc taccgttttg agagactgac tcgggacaaa 2641 tacgcctaca ccaagcagaa agcgacaggg atgaagtaca acttgccttc ctggtgtggc 2701 cgagtcctct ggaagtctta tcccctggtg cacgtggtgt gtcagtctta tggcagtacc 2761 agcgacatca tgacgagtga ccacagccct gtctttgcca catttgaggc aggagtcact 2821 tcccagtttg tctccaagaa cggtcccggg actgttgaca gccaaggaca gattgagttt 2881 ctcaggtgct atgccacatt gaagaccaag tcccagacca aattctacct ggagttccac 2941 tcgagctgct tggagagttt tgtcaagagt caggaaggag aaaatgaaga aggaagtgag 3001 ggggagctgg tggtgaagtt tggtgagact cttccaaagc tgaagcccat tatctctgac 3061 cctgagtacc tgctagacca gcacatcctc atcagcatca agtcctctga cagcgacgaa 3121 tcctatggcg agggctgcat tgcccttcgg ttagaggcca cagaaacgca gctgcccatc 3181 tacacgcctc tcacccacca tggggagttg acaggccact tccaggggga gatcaagctg 3241 cagacctctc agggcaagac gagggagaag ctctatgact ttgtgaagac ggagcgtgat 3301 gaatccagtg ggccaaagac cctgaagagc ctcaccagcc acgaccccat gaagcagtgg 3361 gaagtcacta gcagggcccc tccgtgcagt ggctccagca tcactgaaat catcaacccc 3421 aactacatgg gagtggggcc ctttgggcca ccaatgcccc tgcacgtgaa gcagaccttg 3481 tcccctgacc agcagcccac agcctggagc tacgaccagc cgcccaagga ctccccgctg 3541 gggccctgca ggggagaaag tcctccgaca cctcccggcc agccgcccat atcacccaag 3601 aagtttttac cctcaacagc aaaccggggt ctccctccca ggacacagga gtcaaggccc 3661 agtgacctgg ggaagaacgc aggggacacg ctgcctcagg aggacctgcc gctgacgaag 3721 cccgagatgt ttgagaaccc cctgtatggg tccctgagtt ccttccctaa gcctgctccc 3781 aggaaggacc aggaatcccc caaaatgccg cggaaggaac ccccgccctg cccggaaccc 3841 ggcatcttgt cgcccagcat cgtgctcacc aaagcccagg aggctgatcg cggcgagggg 3901 cccggcaagc aggtgcccgc gccccggctg cgctccttca cgtgctcatc ctctgccgag 3961 ggcagggcgg ccggcgggga caagagccaa gggaagccca agaccccggt cagctcccag 4021 gccccggtgc cggccaagag gcccatcaag ccttccagat cggaaatcaa ccagcagacc 4081 ccgcccaccc cgacgccgcg gccgccgctg ccagtcaaga gcccggcggt gctgcacctc 4141 cagcactcca agggccgcga ctaccgcgac aacaccgagc tcccgcatca cggcaagcac 4201 cggccggagg aggggccacc agggcctcta ggcaggactg ccatgcagaa gcttcgaatt 4261 ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg ccaccatggt gagcaagggc 4321 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 4381 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 4441 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 4501 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc 4561 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 4621 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 4681 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 4741 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac 4801 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag 4861 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 4921 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 4981 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct 5041 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac 5101 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg 5161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt 5281 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 5341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 5401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 5461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca 5521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga 5581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa 5641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc 5701 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc 5761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 5821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc 5881 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 5941 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 6001 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 6061 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 6121 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 6181 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga 6241 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 6301 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 6361 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 6421 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 6481 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 6541 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 6601 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 6661 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 6721 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 6781 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 6841 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 6901 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 6961 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 7021 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 7081 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 7141 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 7201 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac 7261 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt 7321 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga 7381 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt 7441 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag 7501 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 7561 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 7621 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 7681 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 7741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 7801 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 7861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 7921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 7981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 8041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 8101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 8161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 8221 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcct //