LOCUS       sequence-270812-        6713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minigene reporter for alternative splicing analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6713)
  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 6713)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1101/gr.147546.112"; journalName: "Genome Res"; date: "2013-06";
            volume: "23"; issue: "6"; pages: "998-1007"
COMMENT     Sequence Label: pflareA-psd95-exon18.
FEATURES             Location/Qualifiers
     source          1..6713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1783..1792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1928..1930,1931..2644)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHLKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGTDELYK"
     primer_bind     complement(1970..1991)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2231..2250)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      2651..2660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2657..3367
                     /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(2802..2820)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3070..3089
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3489..3610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3526..3545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3580..3599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3617..4072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3754..3775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3966..3985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4099..4203
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4205..4562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4205..4396
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4228..4248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4413..4548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4597..5391
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4597..5391
                     /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(4651..4670)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5261..5280
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5579..5598)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5623..5670
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5999..6587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6488..6507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat
      601 ctaccattgg tgcacctgac tcctgaggag aagtctgccg ttactgccct gtggggcaag
      661 gtgaacgtgg aagagttggt ggtgaggccc tgggccacca gtaagtatca aggttacaag
      721 acaggtttaa ggagaccaat agaaactggg catgtggaga cagagaagac tcacgcgttt
      781 ctgaattgat ctgtctgtcc tctgtccgtc ctcccctcct cccccctcta tcctaattct
      841 gttatcttct cccttcactg tctttgcccg cctgcccggc tctgcccgct gcatcccacc
      901 tctaccaccc ctcttgcctt tgctccactt tcacctttcg ggtcctctcc cccatctcct
      961 acactctgtt ttgccccctt cctgccctcc tgtgcttggc cttccgtctt tctgtccccc
     1021 ctcctcccta ttggtcactc tgcctacatc cctctttctc ttcttttgcc tacatgtccc
     1081 ttggcatctg tcactttccc tttggccaac ctcctgcccc caccttccta tattctctgt
     1141 ctcttcctgg cacctctgtc tgtccttcct gctacctgct ggcccccctc cgcccaccct
     1201 gcctgccccc aggggaagca ctgcatcctt gttgtctcag ccattgccgt gcggcggctg
     1261 caggcggccc acctgcaccc tatcgccatc ttcatccgtc cccgctccct ggagattgtg
     1321 ctgtgagtat ggtcccaggg gctccccctc agctccctcc acctgtgacc ctgggctctc
     1381 aaggtcccct accttgatct gtcctttcag aacctgggac agcaactccc acagtgttca
     1441 aagagctgca ttgagagttg agagtcaggg cttgaaacaa tggggggggg gctttccccc
     1501 ttctcctgcc ttgggttagg caggcctaca gttctcagga gcccccctgg ggcatacaca
     1561 ggctgttgca aaccagaacc tcagaggcaa ctaactggga gtcagtctga attctttcta
     1621 tggggctaaa ggctgaaagg atgagtttca gaggtcctgg atggggtggg ttgccgtgag
     1681 cacatctgta gccccagagg gtatcagaaa tgatatcgtt tgctgtgtgc tccaactatg
     1741 aaagaagttg taatgcagcc tgccaggatc catagtcgac caccatggtg gcttagatcc
     1801 gggcatgtgg agacagagaa gactgttgag tttgtgataa gcactgactc tctctgccta
     1861 ttggtctatt ttccctccct cagtggcggt cgaggacaaa ctctacgcgg acttgccagt
     1921 acgcttcgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1981 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgacgccac
     2041 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2101 caccctcgtg accaccctga cctacggcgt gcagtgcttt agccgctacc ccgaccacct
     2161 gaagcagcac gacttcttca agagtgcaat gcccgaaggc tacgtccagg agcgcaccat
     2221 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2281 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2341 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     2401 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2461 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2521 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcatat
     2581 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcaccgacg agctgtacaa
     2641 gtaaccggtc gccaccatgg tgagcaaggg cgaggagaat aacatggcca tcatcaagga
     2701 gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga
     2761 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2821 gggtggcccc ctgcccttcg cctgggacat cctaaccccc aacttcacct acggctccaa
     2881 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     2941 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3001 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3061 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     3121 gtaccccgag gacggcgccc tgaagggcga gatcaagatg aggctgaagc tgaaggacgg
     3181 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3241 cggcgcctac atcgtcggca tcaagttgga catcacctcc cacaacgagg actacaccat
     3301 cgtggaactg tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3361 taagtaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     3421 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3481 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3541 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3601 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3661 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3721 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3781 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3841 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3901 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3961 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4021 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     4081 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4141 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4201 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4321 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4381 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4441 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4501 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4561 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4621 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4681 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4741 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4801 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4861 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4921 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4981 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5041 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5101 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     5161 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5221 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5281 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5341 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5401 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5461 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5521 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5581 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5641 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5701 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5761 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5821 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5881 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5941 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6121 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6661 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta ata
//
