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" regulatory 539..548 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(726..747) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(987..1006) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" regulatory 1407..1416 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1413..2123 /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(1558..1576) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1826..1845 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2245..2366 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2282..2301) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2336..2355 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2373..2828) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2510..2531) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2722..2741 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2855..2959 /gene="bla" /label=AmpR promoter promoter 2961..3318 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2984..3004) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3169..3304 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3231..3250 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3353..4147 /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(3407..3426) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4017..4036 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4335..4354) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4379..4426 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4755..5343 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5244..5263 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 5518..5821 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5822..6025 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5976..5996 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 tctcagccaa tgccgtgcgg cggctgcagg cggcccacct gcaccctatc gccatcttca 61 tccgtccccg ctccctggag aatgtgctgt gagtatggtc ccaggggctc cccctcagct 121 ccctccacct gtgaccctgg gctctcaagg tcccctacct tgatctgtcc tttcagaacc 181 tgggacagca actcccacag tgttcaaaga gctgcattga gagttgagag tcagggcttg 241 aaacaatggg gggggggctt tcccccttct cctgccttgg gttaggcagg cctacagttc 301 tcaggagccc ccctggggca tacacaggct gttgcaaacc agaacctcag aggcaactaa 361 ctgggagtca gtctgaattc tttctatggg gctaaaggct gaaaggatga gtttcagagg 421 tcctggatgg ggtgggttgc cgtgagcaca tctgtagccc cagagggtat cagaaatgat 481 atcgtttgct gtgtgctcca actatgaaag aagttgtaat gcagcctgcc aggatccacc 541 atggtggctt agatccgggc atgtggagac agagaagact gttgagtttg tgataagcac 601 tgactctctc tgcctattgg tctattttcc ctccctcagt ggcggtcgag gacaaactct 661 acgcggactt gccagtacgc ttcgtgagca agggcgagga gctgttcacc ggggtggtgc 721 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 781 gcgagggcga cgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 841 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgctttagcc 901 gctaccccga ccacctgaag cagcacgact tcttcaagag tgcaatgccc gaaggctacg 961 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 1021 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 1081 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 1141 ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 1201 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 1261 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 1321 agaagcgcga tcatatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 1381 ccgacgagct gtacaagtaa ccggtcgcca ccatggtgag caagggcgag gagaataaca 1441 tggccatcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc 1501 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca 1561 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatccta acccccaact 1621 tcacctacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc 1681 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg 1741 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc 1801 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg 1861 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc 1921 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga 1981 agcccgtgca gctgcccggc gcctacatcg tcggcatcaa gttggacatc acctcccaca 2041 acgaggacta caccatcgtg gaactgtacg aacgcgccga gggccgccac tccaccggcg 2101 gcatggacga gctgtataag taagcggccg cgactctaga tcataatcag ccataccaca 2161 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat 2221 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa 2281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 2341 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa 2401 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 2461 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 2521 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 2581 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg 2641 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc 2701 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc 2761 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca 2821 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 2881 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 2941 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta 3001 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3061 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3121 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3181 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3241 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3301 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga 3361 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 3421 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 3481 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga 3541 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 3601 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 3661 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 3721 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 3781 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 3841 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga 3901 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 3961 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 4021 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 4081 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4141 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 4201 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4261 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct 4321 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg 4381 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt 4441 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 4501 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 4561 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt 4621 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 4681 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 4741 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 4801 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 4861 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 4921 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 4981 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 5041 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 5101 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 5161 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 5221 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 5281 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 5341 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 5401 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag 5461 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt 5521 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 5581 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg 5641 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 5701 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat 5761 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat 5821 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt 5881 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga 5941 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg 6001 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg 6061 gactcagatc taccattggt gcacctgact cctgaggaga agtctgccgt tactgccctg 6121 tggggcaagg tgaacgtgga agagttggtg gtgaggccct gggccacagg taagtatcaa 6181 ggttacaaga caggtttaag gagaccaata gaaactgggc atgtggagac agagaagact 6241 cttgggtttc tgaattgatc tgtctgtcct ctgtccgtcc tcccctcctc ccccctctat 6301 cctaattctg ttatcttctc ccttcactgt ctttgcccgc ctgcccggct ctgcccgctg 6361 catcccacct ctaccacccc tcttgccttt gctccacttt cacctttcgg gtcctctccc 6421 ccatctccta cactctgttt tgcccccttc ctgccctcct gtgcttggcc ttccgtcttt 6481 ctgtcccccc tcctccctat tggtcactct gcctacatcc ctctttctct tcttttgcct 6541 acatgtccct tggcatctgt cactttccct ttggccaacc tcctgccccc accttcctat 6601 attctctgtc tcttcctggc acctctgtct gtccttcctg ctacctgctg gcccccctcc 6661 gcccaccctg cctgccccca ggggaagcac tgcatccttg atg //