LOCUS       sequence-424742-        6287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  minigene construct for Plekha5 E10 alternative splicing .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6287)
  AUTHORS   Zhang M, Ergin V, Lin L, Stork C, Chen L, Zheng S
  TITLE     Axonogenesis Is Coordinated by Neuron-Specific Alternative Splicing 
            Programming and Splicing Regulator PTBP2.
  JOURNAL   Neuron. 2019 Feb 20;101(4):690-706.e10. doi: 
            10.1016/j.neuron.2019.01.022. Epub 2019 Feb 4.
  PUBMED    30733148
REFERENCE   2  (bases 1 to 6287)
  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: 
            "10.1016/j.neuron.2019.01.022"; journalName: "Neuron"; date: 
            "2019-02-20- 20"; volume: "101"; issue: "4"; pages: "690-706.e10"
COMMENT     Sequence Label: pFlare9A-Plekha5E10 WT
FEATURES             Location/Qualifiers
     source          1..6287
                     /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      1369..1378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1514..1516,1517..2230)
                     /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(1556..1577)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1817..1836)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      2237..2246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2243..2953
                     /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(2388..2406)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2656..2675
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3075..3196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3112..3131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3166..3185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3203..3658)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3340..3361)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3552..3571
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3685..3789
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3791..4148
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3791..3982
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3814..3834)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3999..4134
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4183..4977
                     /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             4183..4977
                     /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(4237..4256)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4847..4866
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5165..5184)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5209..5256
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5585..6173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6074..6093
                     /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 ttctgaattc gccttcaaat gcctttcatc atgcatgttc tctcgccgcc
      841 caggccggcc ctcctgcccg cctggcctgg ggtgctgggt caggtgacga ccccacctgc
      901 actcctgtcc cctccttctc ccgtcatttt cactgtcact actctgggtg actctagaat
      961 cctcacattc aaactgaatg ttctttcttc ttacagaatt acctttaatt tccggtgagt
     1021 acatcttttt attaccttaa agctacatag atttatgtgc ttcatggaag agcttgggtt
     1081 ggtttttttg tttgtttgtt tgtttttgtt ttttcgtttt attttggttt tgtttttaaa
     1141 gatcccaaat catagaattt aatttcatgc agttaatttc attgtgtttt tatattgtat
     1201 tggtatttgg gaagtaaata aataaacaat aagaaattct ggtgagcttt atttttatta
     1261 atattatctg gttaattcca attattaata attccagaaa atttaaataa tttctaaatt
     1321 agctacttct tgacgagttg tgcctttcct tggatccata gtcgaccacc atggtggctt
     1381 agatccgggc atgtggagac agagaagact gttgagtttg tgataagcac tgactctctc
     1441 tgcctattgg tctattttcc ctccctcagt ggcggtcgag gacaaactct acgcggactt
     1501 gccagtacgc ttcgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1561 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1621 cgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1681 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgctttagcc gctaccccga
     1741 ccacctgaag cagcacgact tcttcaagag tgcaatgccc gaaggctacg tccaggagcg
     1801 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1861 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1921 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa
     1981 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2041 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2101 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2161 tcatatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca ccgacgagct
     2221 gtacaagtaa ccggtcgcca ccatggtgag caagggcgag gagaataaca tggccatcat
     2281 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
     2341 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2401 gaccaagggt ggccccctgc ccttcgcctg ggacatccta acccccaact tcacctacgg
     2461 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     2521 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     2581 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     2641 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     2701 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa
     2761 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     2821 gctgcccggc gcctacatcg tcggcatcaa gttggacatc acctcccaca acgaggacta
     2881 caccatcgtg gaactgtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     2941 gctgtataag taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3001 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3061 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3121 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3181 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3241 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3301 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3361 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3421 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3481 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3541 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3601 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3661 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3721 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3781 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3961 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4081 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4141 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4681 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4981 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5041 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5101 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5161 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5221 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5281 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5341 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5401 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5461 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5521 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5581 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5641 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5701 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5761 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5821 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5881 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5941 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6001 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6061 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6121 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6181 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6241 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
