LOCUS       sequence-194017-        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        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      1795..1804
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1940..1942,1943..2656)
                     /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(1982..2003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2243..2262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      2663..2672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2669..3379
                     /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(2814..2832)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3082..3101
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3501..3622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3538..3557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3592..3611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3629..4084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3766..3787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3978..3997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4111..4215
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4217..4574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4217..4408
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4240..4260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4425..4560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4609..5403
                     /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             4609..5403
                     /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(4663..4682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5273..5292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5591..5610)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5635..5682
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6011..6599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6500..6519
                     /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 cagtaagtat
      721 caaggttaca agacaggttt aaggagacca atagaaactg ggcatgtgga gacagagaag
      781 actcacgcgt 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 ttgttgtctc agccattgcc
     1261 gtgcggcggc tgcaggcggc ccacctgcac cctatcgcca tcttcatccg tccccgctcc
     1321 ctggagattg 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 tccatagtcg accaccatgg
     1801 tggcttagat ccgggcatgt ggagacagag aagactgttg agtttgtgat aagcactgac
     1861 tctctctgcc tattggtcta ttttccctcc ctcagtggcg gtcgaggaca aactctacgc
     1921 ggacttgcca gtacgcttcg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1981 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2041 gggcgacgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2101 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct ttagccgcta
     2161 ccccgaccac ctgaagcagc acgacttctt caagagtgca atgcccgaag gctacgtcca
     2221 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2281 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2341 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
     2401 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2461 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2521 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2581 gcgcgatcat atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcaccga
     2641 cgagctgtac aagtaaccgg tcgccaccat ggtgagcaag ggcgaggaga ataacatggc
     2701 catcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     2761 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2821 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctaaccc ccaacttcac
     2881 ctacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2941 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     3001 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     3061 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     3121 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaaga tgaggctgaa
     3181 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     3241 cgtgcagctg cccggcgcct acatcgtcgg catcaagttg gacatcacct cccacaacga
     3301 ggactacacc atcgtggaac tgtacgaacg cgccgagggc cgccactcca ccggcggcat
     3361 ggacgagctg tataagtaag cggccgcgac tctagatcat aatcagccat accacatttg
     3421 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3481 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3541 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3601 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3661 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3721 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3781 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3841 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3901 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3961 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4021 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4081 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4141 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4201 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4261 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4321 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4381 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4441 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4501 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4561 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4621 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4681 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4741 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4801 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4861 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4921 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4981 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5041 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5101 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     5161 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5221 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5281 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5341 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5401 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5461 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5521 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5581 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5641 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5701 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5761 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5821 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5881 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5941 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6001 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6061 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6121 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
