LOCUS Exported 6703 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Minigene reporter for alternative splicing analysis. ACCESSION . VERSION . KEYWORDS pflareG-psd95-exon18 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6703) AUTHORS Zheng S, Damoiseaux R, Chen L, Black DL TITLE A broadly applicable high-throughput screening strategy identifies new regulators of Dlg4 (Psd-95) alternative splicing. JOURNAL Genome Res. 2013 Jun;23(6):998-1007. doi: 10.1101/gr.147546.112. Epub 2013 May 1. PUBMED 23636947 REFERENCE 2 (bases 1 to 6703) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6703 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1785..1794 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(1972..1993) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2233..2252) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" regulatory 2653..2662 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2659..3369 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein" /label=mStrawberry /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERA EGRHSTGGMDELYK" primer_bind complement(2804..2822) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3072..3091 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 3491..3612 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3528..3547) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3582..3601 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3619..4074) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3756..3777) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3968..3987 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4101..4205 /gene="bla" /label=AmpR promoter promoter 4207..4564 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4230..4250) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4415..4550 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4477..4496 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4599..5393 /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(4653..4672) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5263..5282 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5581..5600) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5625..5672 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6001..6589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6490..6509 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctaccatt ggtgcacctg actcctgagg agaagtctgc cgttactgcc 661 ctgtggggca aggtgaacgt ggaagagttg gtggtgaggc cctgggccac aggtaagtat 721 caaggttaca agacaggttt aaggagacca atagaaactg ggcatgtgga gacagagaag 781 actcttgggt ttctgaattg atctgtctgt cctctgtccg tcctcccctc ctcccccctc 841 tatcctaatt ctgttatctt ctcccttcac tgtctttgcc cgcctgcccg gctctgcccg 901 ctgcatccca cctctaccac ccctcttgcc tttgctccac tttcaccttt cgggtcctct 961 cccccatctc ctacactctg ttttgccccc ttcctgccct cctgtgcttg gccttccgtc 1021 tttctgtccc ccctcctccc tattggtcac tctgcctaca tccctctttc tcttcttttg 1081 cctacatgtc ccttggcatc tgtcactttc cctttggcca acctcctgcc cccaccttcc 1141 tatattctct gtctcttcct ggcacctctg tctgtccttc ctgctacctg ctggcccccc 1201 tccgcccacc ctgcctgccc ccaggggaag cactgcatcc ttgatgtctc agccaatgcc 1261 gtgcggcggc tgcaggcggc ccacctgcac cctatcgcca tcttcatccg tccccgctcc 1321 ctggagaatg tgctgtgagt atggtcccag gggctccccc tcagctccct ccacctgtga 1381 ccctgggctc tcaaggtccc ctaccttgat ctgtcctttc agaacctggg acagcaactc 1441 ccacagtgtt caaagagctg cattgagagt tgagagtcag ggcttgaaac aatggggggg 1501 gggctttccc ccttctcctg ccttgggtta ggcaggccta cagttctcag gagcccccct 1561 ggggcataca caggctgttg caaaccagaa cctcagaggc aactaactgg gagtcagtct 1621 gaattctttc tatggggcta aaggctgaaa ggatgagttt cagaggtcct ggatggggtg 1681 ggttgccgtg agcacatctg tagccccaga gggtatcaga aatgatatcg tttgctgtgt 1741 gctccaacta tgaaagaagt tgtaatgcag cctgccagga tccaccatgg tggcttagat 1801 ccgggcatgt ggagacagag aagactgttg agtttgtgat aagcactgac tctctctgcc 1861 tattggtcta ttttccctcc ctcagtggcg gtcgaggaca aactctacgc ggacttgcca 1921 gtacgcttcg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag 1981 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgacgcc 2041 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg 2101 cccaccctcg tgaccaccct gacctacggc gtgcagtgct ttagccgcta ccccgaccac 2161 ctgaagcagc acgacttctt caagagtgca atgcccgaag gctacgtcca ggagcgcacc 2221 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac 2281 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg 2341 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag 2401 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag 2461 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 2521 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcat 2581 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcaccga cgagctgtac 2641 aagtaaccgg tcgccaccat ggtgagcaag ggcgaggaga ataacatggc catcatcaag 2701 gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga gttcgagatc 2761 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc 2821 aagggtggcc ccctgccctt cgcctgggac atcctaaccc ccaacttcac ctacggctcc 2881 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag 2941 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag 3001 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc 3061 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg 3121 atgtaccccg aggacggcgc cctgaagggc gagatcaaga tgaggctgaa gctgaaggac 3181 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg 3241 cccggcgcct acatcgtcgg catcaagttg gacatcacct cccacaacga ggactacacc 3301 atcgtggaac tgtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg 3361 tataagtaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt 3421 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat 3481 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 3541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 3601 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt 3661 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat 3721 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat 3781 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac 3841 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc 3901 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga 3961 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca 4021 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg 4081 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 4141 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 4201 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc 4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 4321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 4381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 4441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 4501 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 4561 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 4621 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 4681 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 4741 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat 4801 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 4861 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 4921 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 4981 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 5041 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 5101 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc 5161 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 5221 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 5281 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 5341 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 5401 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 5461 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 5521 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac 5581 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca 5641 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg 5701 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt 5761 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc 5821 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca 5881 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 5941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 6181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6661 gttatcccct gattctgtgg ataaccgtat taccgccatg cat //